Polypeptides and nucleic acids encoding same

ABSTRACT

The present invention provides novel isolated MEMX polynucleotides and polypeptides encoded by the MEMX polynucleotides. Also provided are the antibodies that immunospecifically bind to a MEMX polypeptide or any derivative, variant, mutant or fragment of the MEMX polypeptide, polynucleotide or antibody. The invention additionally provides methods in which the MEMX polypeptide, polynucleotide and antibody are utilized in the detection and treatment of a broad range of pathological states, as well as to other uses.

RELATED APPLICATIONS

[0001] This Application claims priority to U.S. Ser. No. 60/170,564,filed Dec. 14, 1999; U.S. Ser. No. 60/173,165, filed Dec. 27, 1999; U.S.Ser. No. 60/173,362, filed Dec. 27, 1999; U.S. Ser. No. 60/173,544,filed Dec. 29, 1999; U.S. Ser. No. 60/174,962, filed Jan. 5, 2000; U.S.Ser. No. 60/223,929, filed Aug. 9, 2000, and U.S. Ser. No. ______, filedDec. 13, 2000. The contents of these applications are incorporatedherein by reference in their entireties.

FIELD OF THE INVENTION

[0002] The invention generally relates to nucleic acids and polypeptidesencoded therefrom. More specifically, the invention relates to nucleicacids encoding membrane bound and secreted polypeptides, as well asvectors, host cells, antibodies, and recombinant methods for producingthese nucleic acids and polypeptides.

BACKGROUND OF THE INVENTION

[0003] Seven-Pass Transmembrane Receptor

[0004] Seven-pass transmembrane proteins are transmembranal proteinswith seven helices, comprising mostly hydrophobic residues, which serveto facilitate membrane anchoring (see, e.g., Müller, 2000. Curr. Med.Chem. 7: 861-888) and are believed to accommodate the binding site forlow-molecular weight ligands.

[0005] The most well-characterized of the seven-pass transmembranereceptor proteins are the guanine nucleotide-binding signal-transducingprotein (G-protein)-coupled receptors (GPCR) which transduce chemicalsignals through the cytoplasmic membrane by the activation ofintracellular G-proteins. See, e.g., Watson and Arkinstall, THEG-PROTEIN LINKED RECEPTORS, Academic Press, San Diego, Calif., 1994, pp.1-294. In addition, GPCRs constitute the most prominent family ofvalidated drug targets within biomedical research, as approximately 60%of all approved drugs elicit their therapeutic effects by selectivelyinteracting members of this family of proteins and serve as keymolecular targets for therapeutic intervention in a host of diseasestates.

[0006] GPCRs transduce extracellular signals that modulate the activityof a wide variety of biological processes, such as neurotransmission,chemoattraction, cardiac function, olfaction, and vision. Hundreds ofGPCRs signal through one or more of these G proteins in response to alarge variety of stimuli including photons, neurotransmitters, andhormones of variable molecular structure. GPCRs function as a diversefamily of regulatory GTPases which mediate their intracellular actionsthrough the activation of guanine nucleotide-binding signal-transducingproteins (G proteins), that, in the GTP-bound state, bind and activatedownstream membrane-localized effectors. The mechanisms by which theseligands provoke activation of the receptor/G-protein system are highlycomplex and multifactorial. Prominent members of GPCRs include, e.g.,angiotensin II, CCK/gastrin, interleukin 8, endothelin, and the like.See, e.g., van Neuren, 1999. J Recept. Signal Transduct. Res. 9:341-353.

[0007] The GPCR superfamily is evolutionarily conserved and structurallycharacterized by its possessing putative seven-transmembrane (TM)domains with an extracellular amino-terminus and a cytoplasmiccarboxyl-terminus. GPCRs are composed of several independent foldingunits, with the transmembrane domains arranged in a barrel-likestructure with a tightly packed core. The universal adoption of theconserved seven-TM structure by GPCRs, which consequently confers threeintracellular and three extracellular loops along with a TM core,generally is speculated as the minimum necessity to achieve theirstructural stability and functional diversity. None of nearly 2,000GPCRs identified in prokaryotes and eukaryotes to date are known tocontain fewer than seven TM domains.

[0008] In a recent study, alignment of the primary sequencesdemonstrated a high degree of homology within the GPCR transmembraneregions. Three-dimensional (3D) models of 39 GPCRs were generated usingthe refined model of bacteriorhodopsin as a template. Five cationicneurotransmitter receptors (i.e., serotonergic 5-HT2, dopaminergic D2,muscarinic m2, adrenergic alpha 2, and beta 2 receptors) were taken asprototypes and studied in detail. The 3D models of the cationicneurotransmitter receptors, together with their primary structurecomparison, indicate that the agonist binding site is located near theextracellular face of the receptor and involves residues of themembrane-spanning helices 3, 4, 5, 6, and 7. The binding site consistsof a negatively-charged Asp located at the middle of transmembrane helix3 and a hydrophobic pocket containing conserved aromatic residues onhelices 4, 5, 6, and 7. In addition, all the GPCRs were shown to possessinvariant hinge residues, which are thought to be responsible for aconformational change during agonist binding and therefore influencedissociation and association of G-proteins to the receptors. Modulationof the coupling of the G-protein is due to conformation changes withinthis region via hydrophobic interactions and hydrogen bonding. Theinformation of an extracellularly occurring receptor-ligand recognitionevent is transferred through conformational rearrangements within thetransmembranal portion of GPCR to the intracellular compartment. Thus,GPCRs establish a functional and unidirectional link between theexterior of a cell and its cytoplasm.

[0009] Generally, GPCR activation is followed rapidly by a loss ofresponsiveness, termed desensitization, which is then followed by aperiod of recovery or resensitization. These changes in signalingpotential are tightly regulated, primarily via mechanisms that involveGPCR phosphorylation and trafficking to distinct locations within thecell.

[0010] Glutamate and Aspartate Receptors

[0011] Glutamate and Aspartate receptors abound in the Central NervousSystem (CNS), eliciting responses both by ionotropic and metabotropicresponses. Included within the metabotrophic response class areglutamate receptors; which are generally comprised of seven,single-chain transmembrane-spanning proteins. Many cDNAs encodingmetabotropic receptors, as well as ionotropic receptors forN-methyl-D-aspartate, have been identified in recent years. Thediversity of receptor types has also been found to markedly increase asa result of alternative splicing processes and even by single-baseediting of mRNAs. See, e.g., Gilman and Goodman's The PharmacologicalBasis of Therapeutics, Ninth Ed., Hardman, J G, et al. (eds.)McGraw-Hill, New York, 1996, pages 278-282.

[0012] Recently there has been interest in investigating the role ofglutamate receptors in the pathophysiology of schizophrenia. Indeed, thehyperdopaminergic theory of schizophrenia can explain only the positivesymptoms of schizophrenia, whereas the glutamate hypothesis may providea more comprehensive view of the illness. Noorbala, et al. (Piracetam inthe treatment of schizophrenia: implications for the glutamatehypothesis of schizophrenia. PMID: 10583700) undertook a trial toinvestigate whether the combination of haloperidol with piracetam, anootropic agent that modulates the glutamate receptor positively, wasmore effective than haloperidol alone in treating the disease. Theyexamined thirty patients who met the DSM IV criteria for schizophrenia.Patients were allocated in a random fashion, 14 received bothhaloperidol (30 mg/day) and piracetam (3200 mg/day), and 16 patientsreceived only haloperidol (30 mg/day) plus placebo. It was found thatboth protocols significantly decreased the score of the positivesymptoms, the negative symptoms, the general psychopathological symptomsand the total score of PANSS scale over the trial period. Nevertheless,these workers also demonstrated that the combination of haloperidol andpiracetam showed a significant superiority over haloperidol alone in thetreatment of schizophrenic patients. They concluded that piracetam, amember of the nootropic class of drugs and a positive modulator of theglutamate receptor, may be of therapeutic benefit in treatingschizophrenic patients in combination with typical neuroleptic agents.

[0013] Excessive activity of excitatory amino acids released after headtrauma has also been demonstrated to contribute to progressive injury inanimal models and human studies. See, e.g., Morris, et al., 1999. JNeurosurg 91(5): 737-743. Several pharmacological agents that act asantagonists to the glutamate receptor have shown promise in limitingthis progression. The efficacy of the N-methyl-D-aspartate receptorantagonist Selfotel (CGS 19755) was evaluated in two parallel studies ofseverely head injured patients, defined as patients with postresuscitation Glasgow Coma Scale scores of 4 to 8. The Selfotel trialwas terminated prior to completion, however, because of severe adverseeffects on some of the subjects. The results of this trial demonstratethe need for a better understanding of the properties of the glutamatereceptors in the brain, and of the need for discovering more effectiveagonists and antagonists of this receptor.

[0014] Potassium Channel

[0015] The potassium channel mediates the voltage-dependent potassiumion permeability of excitable membranes. Depending upon whether theprotein assumes an opened or closed conformation in response to thevoltage difference across the membrane, the protein forms apotassium-selective channel through which potassium ions may pass inaccordance with their electrochemical gradient.

[0016] The potassium channel has been shown to be an integral membraneprotein. The segment s4 is probably the voltage-sensor and ischaracterized by a series of positively charged amino acids at everythird position. Additionally, the tail may be important in modulation ofchannel activity and/or targeting of the channel to specificsub-cellular compartments. This channel protein belongs to the delayedrectifier class, and to the Shaw potassium channel subfamily.

[0017] IKr (potassium ion channel, rapid response) blockade isineffective in preventing ventricular fibrillation elicited by theinteraction between acute myocardial ischemia and elevated sympatheticactivity. This depends, in-part, upon the fact that adrenergicactivation offsets more than 50% of the action potential prolongingeffect of IKr blockade, and thus impairs its primary mechanism ofaction. The antifibrillatory effect of ersentilide (CK-3579), a novelantiarrhythmic agent which combines blockade of the rapid component ofthe delayed rectifier potassium channel (IKr) with relatively weakbeta-adrenergic blockade, has been examined in a conscious canine modelof lethal arrhythmias. See, Adamson, et al., 1998. Cardiovascular Res.40(1): 56-63). Ersentilide was tested in 19 dogs with a healedmyocardial infarction (MI) undergoing two minutes of circumflex arteryocclusion (CAO) during sub-maximal treadmill exercise. Epicardialmonophasic action potential duration was measured before and afterersentilide in 8 anesthetized open chest dogs at baseline and duringstimulation of the left stellate ganglion at constant paced heart rate.In the control tests 13 of the 19 dogs had ventricular fibrillation (VF)during the exercise and ischemia test, 6 did not. During a subsequentexercise test, ersentilide prevented VF in 820% (11 of 13) of thehigh-risk animals and showed no pro-arrhythmic effects in the 6 dogswithout arrhythmias in the initial test. Ersentilide lowered heart rateat all levels of exercise and during acute myocardial ischemia. Theanti-fibrillatory effect was maintained in 3 of 4 dogs in which heartrate was kept at control levels by atrial pacing. Ersentilide also wasfound to prolonged left ventricular monophasic action potential durationby 30% (from 179+/−6 ms to 233+/−5 ms, p<0.001) at a 360 ms cycle lengthand completely prevented its shortening during sympathetic stimulation.Thus, these authors concluded that the combination of IKr and weakbeta-adrenergic blockade, using ersentilide, represents a very effectiveand safe anti-arrhythmic intervention able to overcome the limitationspresent in drugs devoid of any anti-adrenergic effect. Such acombination may be very useful in the management of post-myocardialinfarction patients at high arrhythmic risk.

[0018] Nair and Grant (1997. Cardiovascular Drugs Ther. 11(2): 149-167)reviewed antiarrhythmic drugs. The goal of developing an antiarrhythmicagent effective against malignant ventricular arrhythmias whilemaintaining a low side-effect profile was evaluated as remainingelusive. In this study, the class III drugs, amiodarone and sotalol,were regarded as the best available agents. However, both drugs possessproperties outside the realm of a pure class III effect, and their useis limited by a variety of dose-related side effects. There are severaldrugs with more selective class III properties currently in development.

[0019] The aforementioned review by Nair and Grant (1997) provides anoverview of the optimal characteristics of an effective theoreticalclass III drug and a summary of the properties of a number of class IIIdrugs under active investigation. An ideal class III antiarrhythmicagent for a reentrant arrhythmia should provide use-dependentprolongation of the action potential duration with slow onset and rapidoffset kinetics. This drug would prolong the effective refractory periodof cardiac tissue selectively at the rapid heart rates achieved duringventricular tachycardia or fibrillation with a delayed onset of action,and a rapid resolution of its effects on resumption of physiologic heartrates. With little effect on the refractory period at normal or slowheart rates, the ability to induce torsade de pointes would be lessened.In contrast to these ideal properties, most currently available andinvestigational agents have a reverse use-dependent effect on the actionpotential duration, producing more effects on the refractory period atslower heart rates. This property results in part from preferentialblock of the rapidly activating component of the delayed rectifierpotassium channel (IKr), with little or no effect on the slowlyactivating component (IKs). The development of a drug with favorableblocking kinetics that selectively blocks IKs may results in lowerproarrhythmic events while still maintaining effective antiarrhythmicproperties.

[0020] Protein Phosphatase I

[0021] Protein phosphatase 1 is believed to act as a scaffold for thelocalization of critical enzymes in glycogen metabolism, includingphosphorylase b, glycogen synthase and phosphorylase kinase. The enzymeis expressed predominantly in insulin-sensitive tissues and was found tomediate the hormonal control of glycogen accumulation in intact cells.

[0022] Hepatic glycogen synthesis is impaired in insulin-dependentdiabetic rats and in adrenalectomized starved rats, and although this isknown to be due to defective activation of glycogen synthase by glycogensynthase phosphatase, the underlying molecular mechanism has not beendelineated. Glycogen synthase phosphatase comprises the catalyticsubunit of protein phosphatase 1 (PP1) complexed with the hepaticglycogen-binding subunit, termed GL. In liver extracts ofinsulin-dependent diabetic and adrenalectomized starved rats, the levelof GL was shown by immunoblotting to be substantially reduced comparedwith that in control extracts, whereas the level of PP1 catalyticsubunit was not affected by these treatments. See, Doherty, et al.,1998. Biochem. J 333: 253-257. Insulin administration to diabetic ratsrestored the level of GL and prolonged administration raised it abovethe control levels, whereas re-feeding partially restored the GL levelin adrenalectomized starved rats. The regulation of GL protein levels byinsulin and starvation/feeding was shown to correlate with changes inthe level of the GL mRNA, indicating that the long-term regulation ofthe hepatic glycogen-associated form of PP 1 by insulin, and hence theactivity of hepatic glycogen synthase, is predominantly mediated throughchanges in the level of the GL mRNA. (PMID: 9657963, UI: 98324884).

[0023] Retinol-Binding Protein

[0024] Retinol-binding protein (RBP) is the specific carrier for retinol(vitamin A1) in the blood. Low RBP level in the blood has been found tobe associated with low serum retinol level in keratomalacia patients.Familial hypo-RB proteinemia has been found to predispose the probandchild to keratomalacia during measles infection, despite good nutrition.See, e.g., Attard-Montalto, et al. described a girl with intermittentorange discoloration of her palms, soles, and face and with carotenemiaassociated with persistently low levels of both vitamin A andserum-specific retinol-binding protein. These authors postulated thatthe low serum retinol-binding protein concentration resulted in the slowuptake and release of vitamin A by the liver. The conversion of caroteneto vitamin A was consequently inhibited and this resulted inhypercarotenemia. Vitamin A supplements were unable to raise the serumvitamin A concentration and did not relieve the carotenemia.

[0025] Seeliger, et al., reported the ocular phenotype in retinoldeficiency due to a hereditary defect in retinol-binding proteinsynthesis. Two affected sisters, aged 17 and 13 years, were compoundheterozygous for missense mutations in the RBP4 gene. Each affectedsister had had night vision problems since early childhood but wasotherwise well. Visual acuities were slightly reduced: 20/40 in the 17year old and 20/25 in the 13 year old. Both affected sibs had nodetectable serum RBP, retinol levels less than 20% of normal, and normalretinyl esters.

[0026] RBP gene has been mapped to the long arm of chromosome 10 and ishomologous to bovine beta-lactoglobulin.

SUMMARY OF THE INVENTION

[0027] The invention is based, in part, upon the discovery of a novelpolynucleotide sequences encoding novel polypeptides. Nucleic acidsencoding these polypeptides, and derivatives and fragments thereof, willhereinafter be collectively designated as “MEMX”.

[0028] Accordingly, in one aspect, the invention provides an isolatednucleic acid molecule that includes the sequence of SEQ ID NO: 1, 3, 57, 9, 11, 13, or 15, or a fragment, homolog, analog or derivativethereof. The nucleic acid can include, e.g., a nucleic acid sequenceencoding a polypeptide at least 80% identical to a polypeptide thatincludes the amino acid sequences of SEQ ID NO: 2, 4, 6, 8, 10, 12, 14,or 16. The nucleic acid can be, e.g., a genomic DNA fragment, or a cDNAmolecule.

[0029] Also included in the invention is a vector containing one or moreof the nucleic acids described herein, and a cell containing the vectorsor nucleic acids described herein.

[0030] The invention is also directed to host cells transformed with avector comprising any of the nucleic acid molecules described above.

[0031] In another aspect, the invention includes a pharmaceuticalcomposition that includes an MEMX nucleic acid and a pharmaceuticallyacceptable carrier or diluent.

[0032] In a further aspect, the invention includes a substantiallypurified MEMX polypeptide, e.g., any of the MEMX polypeptides encoded byan MEMX nucleic acid, and fragments, homologs, analogs, and derivativesthereof. The invention also includes a pharmaceutical composition thatincludes an MEMX polypeptide and a pharmaceutically acceptable carrieror diluent.

[0033] In still a further aspect, the invention provides an antibodythat binds specifically to an MEMX polypeptide. The antibody can be,e.g., a monoclonal or polyclonal antibody, and fragments, homologs,analogs, and derivatives thereof. The invention also includes apharmaceutical composition including MEMX antibody and apharmaceutically acceptable carrier or diluent. The invention is alsodirected to isolated antibodies that bind to an epitope on a polypeptideencoded by any of the nucleic acid molecules described above.

[0034] The invention also includes kits comprising any of thepharmaceutical compositions described above.

[0035] The invention further provides a method for producing an MEMXpolypeptide by providing a cell containing an MEMX nucleic acid, e.g., avector that includes an MEMX nucleic acid, and culturing the cell underconditions sufficient to express the MEMX polypeptide encoded by thenucleic acid. The expressed MEMX polypeptide is then recovered from thecell. Preferably, the cell produces little or no endogenous MEMXpolypeptide. The cell can be, e.g., a prokaryotic cell or eukaryoticcell.

[0036] The invention is also directed to methods of identifying an MEMXpolypeptide or nucleic acid in a sample by contacting the sample with acompound that specifically binds to the polypeptide or nucleic acid, anddetecting complex formation, if present.

[0037] The invention further provides methods of identifying a compoundthat modulates the activity of an MEMX polypeptide by contacting an MEMXpolypeptide with a compound and determining whether the MEMX polypeptideactivity is modified.

[0038] The invention is also directed to compounds that modulate MEMXpolypeptide activity identified by contacting an MEMX polypeptide withthe compound and determining whether the compound modifies activity ofthe MEMX polypeptide, binds to the MEMX polypeptide, or binds to anucleic acid molecule encoding an MEMX polypeptide.

[0039] In another aspect, the invention provides a method of determiningthe presence of or predisposition of an MEMX-associated disorder in asubject. The method includes providing a sample from the subject andmeasuring the amount of MEMX polypeptide in the subject sample. Theamount of MEMX polypeptide in the subject sample is then compared to theamount of MEMX polypeptide in a control sample. An alteration in theamount of MEMX polypeptide in the subject protein sample relative to theamount of MEMX polypeptide in the control protein sample indicates thesubject has a tissue proliferation-associated condition. A controlsample is preferably taken from a matched individual, i.e., anindividual of similar age, sex, or other general condition but who isnot suspected of having a tissue proliferation-associated condition.Alternatively, the control sample may be taken from the subject at atime when the subject is not suspected of having a tissueproliferation-associated disorder. In some embodiments, the MEMX isdetected using an MEMX antibody.

[0040] In a further aspect, the invention provides a method ofdetermining the presence of or predisposition of an MEMX-associateddisorder in a subject. The method includes providing a nucleic acidsample, e.g., RNA or DNA, or both, from the subject and measuring theamount of the MEMX nucleic acid in the subject nucleic acid sample. Theamount of MEMX nucleic acid sample in the subject nucleic acid is thencompared to the amount of an MEMX nucleic acid in a control sample. Analteration in the amount of MEMX nucleic acid in the sample relative tothe amount of MEMX in the control sample indicates the subject has atissue proliferation-associated disorder.

[0041] In a still further aspect, the invention provides a method oftreating or preventing or delaying an MEMX-associated disorder. Themethod includes administering to a subject in which such treatment orprevention or delay is desired an MEMX nucleic acid, an MEMXpolypeptide, or an MEMX antibody in an amount sufficient to treat,prevent, or delay a tissue proliferation-associated disorder in thesubject.

[0042] Unless otherwise defined, all technical and scientific terms usedherein have the same meaning as commonly understood by one of ordinaryskill in the art to which this invention belongs. Although methods andmaterials similar or equivalent to those described herein can be used inthe practice or testing of the present invention, suitable methods andmaterials are described below. All publications, patent applications,patents, and other references mentioned herein are incorporated byreference in their entirety. In the case of conflict, the presentspecification, including definitions, will control. In addition, thematerials, methods, and examples are illustrative only and not intendedto be limiting.

[0043] Other features and advantages of the invention will be apparentfrom the following detailed description and claims.

BRIEF DESCRIPTION OF THE DRAWINGS

[0044]FIG. 1: illustrates the nucleotide sequence of the Seven-PassTransmembrane Receptor-Like Protein of the invention [SEQ ID NO:1]. Thestart and stop codons are shown in bold font.

[0045]FIG. 2: illustrates the amino acid sequence [SEQ ID NO:2] encodedby the coding sequence shown in FIG. 1.

[0046]FIG. 3: illustrates the BLASTN identity searches leading to thenucleic acid sequence [SEQ ID NO:1].

[0047]FIG. 4: illustrates the BLASTX identity search for the amino acidsequence [SEQ ID NO:2].

[0048]FIG. 5: illustrates the BLASTP identity search for the amino acidsequence [SEQ ID NO:2].

[0049]FIG. 6: illustrates the ClustalW alignment of the amino acidsequence [SEQ ID NO:2].

[0050]FIG. 7: illustrates the nucleotide sequence, including thesequence encoding a glutamate receptor variant (21659259 EXT 1) of theinvention [SEQ ID NO:3]. The start and stop codons are shown in boldfont.

[0051]FIG. 8: illustrates the amino acid sequence [SEQ ID NO:4] encodedby the coding sequence of 21659259 EXT 1 shown in FIG. 7.

[0052]FIG. 9: illustrates the BLASTN identity searches leading to thenucleic acid sequence [SEQ ID NO:3] of variant 21659259 EXT 1.

[0053]FIG. 10: illustrates the BLASTX identity search for the amino acidsequence [SEQ ID NO:4] of variant 21659259 EXT 1.

[0054]FIG. 11: illustrates the ClustalW alignment of variant 21659259EXT 1.

[0055]FIG. 12: illustrates the nucleotide sequence, including thesequence encoding a glutamate receptor variant (21659259 EXT 2) of theinvention [SEQ ID NO:5].

[0056]FIG. 13: illustrates the amino acid sequence [SEQ ID NO:6] encodedby the coding sequence of 21659259 EXT 2 of FIG. 12.

[0057]FIG. 14: illustrates the BLASTN identity searches lea ding to thenucleic acid sequence [SEQ ID NO:5] of variant 21659259 EXT 2.

[0058]FIG. 15: illustrates the BLASTX identity search for the amino acidsequence [SEQ ID NO:6] of variant 21659259 EXT 2.

[0059]FIG. 16: illustrates the ClustalW alignment of variant 21659259EXT 2.

[0060]FIG. 17: illustrates the nucleotide sequence, including thesequence encoding a glutamate receptor variant (21659259 EXT 3) of theinvention [SEQ ID NO:7].

[0061]FIG. 18: illustrates the amino acid sequence [SEQ ID NO:8] encodedby the coding sequence of 21659259 EXT 3 of FIG. 17.

[0062]FIG. 19: illustrates the BLASTN identity searches leading to thenucleic acid sequence [SEQ ID NO:7] of variant 21659259 EXT 3.

[0063]FIG. 20: illustrates the BLASTX identity search for the amino acidsequence [SEQ ID NO:8] of variant21659259 EXT 3.

[0064]FIG. 21: illustrates the ClustalW alignment of variant 21659259EXT 3.

[0065]FIG. 22: illustrates the ClustalW alignment of the three splicevariants of the glutamate receptor of the present invention.

[0066]FIG. 23: illustrates the nucleotide sequence [SEQ ID NO:9] of thepotassium channel protein of the invention. A putative untranslatedregion 5′ to the start codon is shown by underlining, whereas the startand termination codons are shown in bold font.

[0067]FIG. 24: illustrates the amino acid sequence [SEQ ID NO: 10]encoded by the coding sequence shown in FIG. 23.

[0068]FIG. 25: illustrates the BLASTX identity search for the protein ofthe invention [SEQ ID NO:10].

[0069]FIG. 26: illustrates the nucleotide sequence including thesequence encoding the phosphatase 1-like protein of the invention [SEQID NO: 11]. Putative untranslated regions 5′ to the start codon and 3′to termination codon are shown by underlining, and the start and stopcodons are shown in bold font.

[0070]FIG. 27: illustrates the amino acid sequence [SEQ ID NO: 12]encoded by the coding sequence shown in FIG. 26.

[0071]FIG. 28: illustrates BLASTN identity search for the nucleic acidencoding the phosphatase 1-like protein of the invention.

[0072]FIG. 29: illustrates the BLASTX identity search for thephosphatase 1-like protein of the invention.

[0073]FIG. 30: illustrates the ClustalW alignment of the phosphatase1-like protein of the invention.

[0074]FIG. 31: illustrates the nucleotide sequence [SEQ ID NO: 13],including the sequence encoding the protein resembling retinol-bindingprotein, of the invention. Putative untranslated regions 5′ to the startcodon and 3′ to the termination codon are shown by underlining, and thestart and stop codons are shown in bold font.

[0075]FIG. 32: illustrates the amino acid sequence [SEQ ID NO: 14]encoded by the coding sequence shown in FIG. 31.

[0076]FIG. 33: illustrates the BLASTX identity search for theretinol-binding-like protein of the invention shown in FIG. 32.

[0077]FIG. 34: illustrates the nucleotide sequence [SEQ ID NO: 15],including the sequence encoding the protein resembling retinol-bindingprotein, of the invention.

[0078]FIG. 35: illustrates the amino acid sequence [SEQ ID NO:16]encoded by the coding sequence shown in FIG. 34.

[0079]FIG. 36: illustrates the BLASTP identity search for theretinol-binding-like protein of the invention shown in FIG. 35.

[0080]FIG. 37: illustrates the ClustalW alignment of theretinol-binding-like protein of the invention shown in FIG. 35.

DETAILED DESCRIPTION OF THE INVENTION

[0081] The present invention provides novel nucleotides and polypeptidesencoded thereby. Included in the invention are the novel nucleic acidsequences and their polypeptides. The sequences are collectivelydesignated as “MEMX nucleic acids” or “MEMX polynucleotides” and thecorresponding encoded polypeptides are referred to as “MEMXpolypeptides” or “MEMX proteins.” Unless indicated otherwise, “MEMX” ismeant to refer to any of the novel sequences disclosed herein. Table 1,below, provides a summary of the MEMX nucleic acids and their encodedpolypeptides. TABLE 1 MEMX SEQ ID Assign- Internal NO:) nu- SEQ ID NO:ment Identification cleic acid) (polypeptide) Homology 1 Construct of 12 Seven-Pass AL021392, Transmembrane AL031588, Receptor Protein andAL031597 2 21659259 3 4 Glutamate Receptor EXT 1 3 21659259 5 6Glutamate Receptor EXT 2 4 21659259 7 8 Glutamate Receptor EXT 3 516418841 9 10  Potassium Channel Protein 6 AC016485_A 11  12 Phosphatase I Protein 7 AC018653_A 13  14  Retinol-Binding Protein 8AC18653A dal 15  16  Retinol-Binding Protein

[0082] MEMX nucleic acids and their encoded polypeptides are useful in avariety of applications and contexts. The various MEMX nucleic acids andpolypeptides according to the invention are useful as members of theprotein families according to the presence of domains and sequencerelatedness to previously described proteins. Additionally, MEMX nucleicacids and polypeptides can also be used to identify proteins that aremembers of the family to which the MEMX polypeptides belong.

[0083] For example, MEMI is homologous to members of the Seven-PassTransmembrane Receptor Protein family of proteins. Thus, the MEMInucleic acids and polypeptides, antibodies and related compoundsaccording to the invention will be useful in therapeutic and diagnosticapplications in immunotherapy, viral infections, neurological disorders(e.g., Alzheimer's disease or Parkinson's disease), cancer (e.g., breastor neuroblastoma), nephrology, and female reproductive health.

[0084] MEM2, MEM3, and MEM4 are homologous to members of the GlutamateReceptor family of proteins. Thus, the MEM2 through MEM4 nucleic acidsand polypeptides, antibodies and related compounds according to theinvention will be useful in therapeutic and diagnostic applicationstargeted to lung and/or brain. In brain, it may serve as a targetreceptor for treating schizophrenia or reducing neuronal damagefollowing head injury.

[0085] MEM5 is homologous to members of the Potassium Channel Proteinfamily of proteins. Thus, the MEM5 nucleic acids and polypeptides,antibodies and related compounds according to the invention will beuseful in therapeutic and diagnostic applications in the treatment ofheart and other muscular disorders (e.g., anti-arrhythmic agents),supplementation of defective clotting Factor XI in clottingdeficiencies, and cobalamin-deficiencies (e.g., pernicious anemia).

[0086] MEM6 is homologous to members of the Phosphatase I Protein familyof proteins. Thus, the MEM6 nucleic acids and polypeptides, antibodiesand related compounds according to the invention will be useful intherapeutic and diagnostic applications in the treatment of diabetes andrelated disorders originating in dysregulation of glycogen metabolism.

[0087] MEM7 and MEM8 are homologous to members of the Retinol-BindingProtein family of proteins. Thus, the MEM7 and MEM8 nucleic acids andpolypeptides, antibodies and related compounds according to theinvention will be useful in therapeutic and diagnostic applications inthe treatment of vision-related disorders (e.g., keratomalacia), andcancer and/or similar neoplastic pathologies.

[0088] The MEMX nucleic acids and polypeptides can also be used toscreen for molecules, which inhibit or enhance MEMX activity orfunction. Additional utilities for MEMX nucleic acids and polypeptidesaccording to the invention are disclosed herein.

[0089] MEM1

[0090] An MEM1 sequence according to the invention iincludes a nucleicacid sequence encoding a polypeptide related to the seven-passtransmembrane receptor family of proteins. The nucleotide sequence [SEQID NO:1] of the novel nucleic acid (designated CuraGen Acc. Nos.AL021392, AL031588, and AL031597) encoding a novel protein resemblingthe seven-pass transmembrane receptor proteins is shown in FIG. 1. AnOpen Reading Frame (ORF) was identified beginning with an atg initiationcodon and ending with a tga termination codon. Putative untranslatedregions upstream from the initiation codon and downstream from thetermination codon are shown by underlining, and the start and stopcodons are shown in bold letters. The amino acid sequence [SEQ ID NO:2]of the encoded protein is presented using the one-letter code in FIG. 2.

[0091] In a BlastN search of nucleic acid sequence databases (see, FIG.3), it was found, e.g., that the MEM1 nucleic acid sequence has 203 of218 bases (93%) positive and 203 of 218 bases (93%) identical tosequence (designated HS1163J1) which contains: the 3′ region of a genefor a novel KIAA0279-lile EGF-like domain containing a protein similarto murine Celsr1 and rat MEGF2; a novel gene for a protein similar to C.elegans B0035.16 and bacterial tRNA(5′-Methylaminomethyl-2-thiouridylate)-Methyltransferases; and the 3′region of a novel gene for a protein similar to murine B99.

[0092] In a search of amino acid databases, the MEM1 protein of theinvention was found to have 172 of 186 amino acid residues (91%)positive with, and 162 of 186 amino acid residues (87%) identical to theseven-pass transmembrane receptor protein precusor MouseA (ptnr:PIR-ID:T14119, see, FIG. 4) which is a member of the Celsr family ofseven-pass transmembrane receptor proteins which are expressed duringembryogenesis in the mouse. In a BlastP search (see, FIG. 5), theprotein of the present invention was found to have 2345 of 2632 aminoacid residues (89%) positive with, and 2139 of 2632 amino acid residues(81%) identical to the amino acid residue seven-pass transmembranereceptor protein precusor MouseA.

[0093] A multiple sequence alignment is illustrated in FIG. 6, with theprotein of the invention being shown on Line 2, in a ClustalW analysiscomparing the protein of the invention with related protein sequences.

[0094] The novel nucleic acid of the invention encoding a proteinresembling the seven-pass transmembrane receptor family of proteinsincludes the nucleic acid whose sequence [SEQ ID NO: 1] is provided inFIG. 1, or a fragment thereof. The invention also includes a mutant orvariant nucleic acid any of whose bases may be changed from thecorresponding base shown in FIG. 1, while still encoding a protein thatmaintains its retinol-binding activities and physiological functions, ora fragment of such a nucleic acid. The invention further includesnucleic acids whose sequences are complementary to those just described,including nucleic acid fragments that are complementary to any of thenucleic acids just described. The invention additionally includesnucleic acids or nucleic acid fragments, or complements thereto, whosestructures include chemical modifications. Such modifications include,by way of non-limiting example, modified bases, and nucleic acids whosesugar phosphate backbones are modified or derivatized. Thesemodifications are carried out at least in part to enhance the chemicalstability of the modified nucleic acid, such that they may be used, forexample, as antisense binding nucleic acids in therapeutic applicationsin a subject. In the mutant or variant nucleic acids, and theircomplements, up to 20% or more of the bases may be so changed.

[0095] The novel protein of the invention includes the proteinsresembling seven-pass transmembrane receptor proteins whose sequence[SEQ ID NO:2] is provided in FIG. 2. The invention also includes amutant or variant protein any of whose residues may be changed from thecorresponding residue shown in FIG. 2, while still encoding a proteinthat maintains its proteins resembling retinol-binding activities andphysiological functions, or a functional fragment thereof. In the mutantor variant protein, up to 20% or more of the residues may be so changed.The invention further encompasses antibodies and antibody fragments,such as F_(ab) or (_(Fab))₂, that bind immunospecifically to any of theproteins of the invention.

[0096] MEM2, MEM3, and MEM4

[0097] An MEM2, MEM3, and MEM4 sequence according to the inventionincludes a nucleic acid sequence encoding a polypeptide related to thehuman glutamate receptor family of proteins. Three variants of a humanglutamate receptor MEM2 (Internal identification No. 21659259 EXT 1);MEM3 (Internal identification No. 21659259 EXT 2); and MEM4 (Internalidentification No. 21659259 EXT 3) are disclosed in the presentinvention. These differing sequences apparently result from splicevariants (or a similar deletion) at the nucleic acid level and resemblea lung-specific, splice-form of a previously reported glutamate receptor(SPTREMBL-ACC:060391). Each of the three splice variants will bediscussed below.

[0098] Splice Variant 21659259 EXT 1 (MEM2)

[0099] The nucleotide sequence of one splice variant of the presentinvention MEM2 (Internal Identification No. 21659259 EXT 1) is shown inFIG. 7 [SEQ ID NO:3]. An Open Reading Frame (ORF) was identifiedbeginning with the atg initiation codon and ending with the tgatermination codon. The start and termination codons are shown in boldletters. The encoded protein is illustrated using one-letter amino acidcode in FIG. 8 [SEQ ID NO:4].

[0100] In this splice variant, the difference was found at amino acidresidue 360, where 73 amino acids residues were shown to be deleted(i.e., “spliced-out”). These amino acid residues are also present in thebest Blast-X protein match (SPTREMBL-ACC:O60391). It is important tonote that these 73 amino acids are also spliced out in a reportedglutamate receptor from human brain (SWISSPROT-ACC:Q14957). Therefore,this variant may represent an isoform of a glutamate receptor that ispresent in both lung and brain.

[0101] BLASTN comparisons leading to the assembly of the 21659259 EXT 1variant of this invention are illustrated in FIG. 9. As noted above, thesequences of the present invention match a genomic sequence(SPTREMBL-ACC:060391). In assembling and verifying the sequences, onecorrection was made to the SeqCalling assembly, which added a G atnucleotide 104 of the assembly. It was noted that the sequencing traceappearance also suggested that another G could be present in thesequence at basepair 104. Furthermore, adding the G corrected a frameshift in the protein and resulted in a better Blast-X match with otherreported glutamate receptors. This gene, 21659259 EXT 1, differs fromthe previously reported gene (SPTREMBL-ACC:O60391). The protein in thepublic database (SPTREMBL-ACC:O60391) includes 73 amino acids that aremissing in the present 21659259 EXT 1 sequence. It is believed that thepresently disclosed assembly (21659259 EXT 1), which is derived fromfetal lung tissue, represents a splice variant of the reported protein.This represents omission of bases 22806-23025 of the genomic sequence(GENBANK-ID:AC004528).

[0102] The protein in the public database (SPTREMBL-ACC:O60391)additionally includes 6 amino acid residues at the beginning of the exon(i.e., basepairs 25855-26000) of the genomic sequence(GENBANK-ID:AC004528). In the presently disclosed sequence, however, thesame exon includes only the region between basepairs 25873-26000 bp, anddoes not contain the 18 nucleotides which lie between basepairs25855-25873 of the genomic sequence. Accordingly, the protein variant21659259 EXT 1 of the present invention lacks the six amino acids,present in the human and rat reference sequences, encoded by thesemissing bases.

[0103] Additionally, the protein found in the public database(SPTREMBL-ACC:O60391) also lacks the last exon containing 430 bppredicted by GenScan in the present invention. This exon terminates withthe stop codon TGA. BLASTX comparisons used in identifying variant21659259 EXT 1 are shown in FIG. 10.

[0104] A multiple sequence alignment of variant 21659259 EXT 1 isillustrated in FIG. 11, with the protein of the invention being shown onLine 3, in a ClustalW analysis comparing the protein of the inventionwith related protein sequences. The 73-residue and 6-residue deletionsare shown, as is the C-terrninal extension.

[0105] Splice Variant 21659259 EXT 2 (MEM3)

[0106] The nucleotide sequence [SEQ ID NO:5] of a second splice variant,MEM3 (Internal Identification No. 21659259 EXT 2), of the presentinvention is shown in FIG. 12. An Open Reading Frame (ORF) wasidentified beginning with an atg initiation codon and ending with a tgatermination codon. The start and termination codons are shown in boldletters. The encoded protein [SEQ ID NO:6] is illustrated using theone-letter amino acid code in FIG. 13.

[0107] Two of the three distinctions found in MEM2 (the 21659259 EXT 1variant) were also demonstrated to be present with this splice variant.However, it was believed that the eighteen nucleotide omission noted forMEM2 (21659259 EXT 1) should be included in view of the fact that thisfragment is present in a variety of glutamate receptors. Thus the aminoacids encoded by these nucleotides are included in the amino acidsequence of this variant.

[0108] BLASTN comparisons leading to the assembly of the 21659259 EXT 2variant of this invention are included in FIG. 14. BLASTX comparisonsused in identifying variant 21659259 EXT 2 are shown in FIG. 15.

[0109] A multiple sequence alignment is of MEM3 variant 21659259 EXT 2given in FIG. 16, with the protein of the invention being shown on Line3, in a ClustalW analysis comparing the protein of the invention withrelated protein sequences. The 73-residue deletion is shown, as is thecarboxyl-terminal extension.

[0110] Splice Variant 21659259 EXT 3 (MEM4)

[0111] The nucleotide sequence [SEQ ID NO:7] of a third splice variant,MEM4 (Internal Identification No. 21659259 EXT 3), of the invention isshown in the nucleotide sequence of FIG. 17. An open reading frame wasidentified beginning with an atg initiation codon and ending with a tgatermination codon. The start and stop codons are in bold letters. Theamino acid sequence [SEQ ID NO:8] of the encoded protein is presentedusing the one-letter code in FIG. 18.

[0112] One of the three distinctions found with MEM2 (21659259 EXT 1)also occur in this variant. Due to the fact that these fragments havebeen shown to be present in a variety of glutamate receptors, both theeighteen nucleotide omission noted for MEM2 (21659259 EXT 1), as well asthe 73 amino acid deletion, were included in the sequence of this splicevariant. Thus, the amino acid sequences represented by these deletionsare included in the amino acid sequence of this variant.

[0113] BLASTN comparisons leading to the assembly of MEM4 areillustrated in FIG. 19; whereas the BLASTX comparisons used inidentifying MEM4 are illustrated FIG. 20. Although the match for 900 ofthe 901 residues of the SPTREMBL-ACC:O60391 sequence is 100% identicalto that of 21659259 EXT 3 , the public protein (SPTREMBL-ACC:O60391) isfound to lack the terminal 143 amino acids included in the splicevariants of the present invention.

[0114] ClustalW analysis comparing variant 21659259 EXT 3 with relatedprotein sequences is illustrated in FIG. 21, with the protein of theinvention being shown on Line 3. In addition, the carboxyl-terminalextension is shown.

[0115] A comparative alignment of the three splice variants of thepresent invention MEM2, MEM3, and MEM4 (i.e., 21659259 EXT 1; 21659259EXT 2; and 21659259 EXT 3) is shown in FIG. 22.

[0116] The novel nucleic acid of the invention encoding a glutamatereceptor includes the nucleic acid whose sequence is provided in FIG. 7[SEQ ID NO:3]; FIG. 12 [SEQ ID NO:5]; and FIG. 17 [SEQ ID NO:7], orfragments thereof. The present invention also includes a mutant orvariant nucleic acid any of whose bases may be changed from thecorresponding base shown in FIGS. 7, 12, and 17, while still encoding aprotein that maintains its glutamate receptor-like activities andphysiological functions, or a fragment of such a nucleic acid. Theinvention further includes nucleic acids whose sequences arecomplementary to those just described, including nucleic acid fragmentsthat are complementary to any of the nucleic acids just described. Theinvention additionally includes nucleic acids or nucleic acid fragments,or complements thereto, whose structures include chemical modifications.Such modifications include, by way of non-limiting example, modifiedbases, and nucleic acids whose sugar phosphate backbones are modified orderivatized. These modifications are carried out at least in part toenhance the chemical stability of the modified nucleic acid, such thatthey may be used, for example, as antisense binding nucleic acids intherapeutic applications in a subject. In the mutant or variant nucleicacids, and their complements, up to 20% or more of the bases may be sochanged.

[0117] The novel protein of the invention includes the followingproteins: FIG. 8 [SEQ ID NO:4]; FIG. 13 [SEQ ID NO:6]; and FIG. 18 [SEQID NO:8]. The invention also includes a mutant or variant protein any ofwhose residues may be changed from the corresponding residue shown inFIG. 8, FIG. 13, and FIG. 18, while still encoding a protein thatmaintains its glutamate receptor-like protein-like activities andphysiological functions, or a functional fragment thereof. In the mutantor variant protein, up to 20% or more of the residues may be so changed.The invention further encompasses antibodies and antibody fragments,such as F_(ab) or (F_(ab))₂, that bind immunospecifically to any of theproteins of the invention.

[0118] MEM5

[0119] An MEM5 sequence according to the invention includes a nucleicacid sequence encoding a polypeptide related to the potassium channelproteins. The novel nucleic acid sequence [SEQ ID NO:9] of 1110nucleotides (Internal Identification No. 16418841_EXT) encoding a ionchannel-like protein is shown in FIG. 23. An Open Reading Frame (ORF) of828 nucleotides was identified beginning with an atg initiation codonand ending with a tga termination codon (see, FIG. 23; [SEQ ID NO:9]).Putative untranslated regions, one upstream from the initiation codonand another downstream of the termination codon, are shown byunderlining in FIG. 23, whereas the start and termination codons areshown in bold letters. The sequence of the encoded protein [SEQ ID NO:10] comprising 275 amino acid residues is presented using the one-letteramino code in FIG. 24.

[0120] In a search of sequence databases (see, FIG. 25), it was found,e.g., that the nucleic acid sequence of the protein of the invention hasfound to have 286 of 286 amino acid residues (100%) identical to, and286 of 286 amino acid residues (100%) positive with, the Human potassiumchannel protein K⁺Hnov42 (patp:Y34130; see, International PublicationNo. WO 9943696 Al).

[0121] A hydrophobicity plot shows that the protein of the invention hasa short, N-terminal, hydrophilic sequence (1-40 aa), followed by ahydrophobic region (41-65 aa, peak hydrophobicity=1), followed by ahydrophilic C-terminus. Although a SignalP analysis suggests that thereis no signal peptide, the hydrophobic region at 41-65 may neverthelessbe a cleavable signal peptide.

[0122] The novel nucleic acid of the invention includes the nucleic acidwhose sequence [SEQ ID NO:9] is provided in FIG. 23, or a fragmentthereof. The invention also includes a mutant or variant nucleic acidany of whose bases may be changed from the corresponding base shown inFIG. 23, while still encoding a protein that maintains its activitiesand physiological functions, or a fragment of such a nucleic acid. Theinvention further includes nucleic acids whose sequences arecomplementary to those just described, including nucleic acid fragmentsthat are complementary to any of the nucleic acids just described. Theinvention additionally includes nucleic acids or nucleic acid fragments,or complements thereto, whose structures include chemical modifications.Such modifications include, by way of non-limiting example, modifiedbases, and nucleic acids whose sugar phosphate backbones are modified orderivatized. These modifications are carried out at least in part toenhance the chemical stability of the modified nucleic acid, such thatthey may be used, for example, as antisense binding nucleic acids intherapeutic applications in a subject. In the mutant or variant nucleicacids, and their complements, up to 20% or more of the bases may be sochanged.

[0123] The novel protein of the invention includes the protein whosesequence [SEQ ID NO:10] is provided in FIG. 24. The invention alsoincludes a mutant or variant protein any of whose residues may bechanged from the corresponding residue shown in FIG. 24, while stillencoding a protein that maintains its potassium channel protein-likeactivities and physiological functions, or a functional fragmentthereof. In the mutant or variant protein, up to 20% or more of theresidues may be so changed. The invention further encompasses antibodiesand antibody fragments, such as F_(ab) or (F_(ab))₂, that bindimmunospecifically to any of the proteins of the invention.

[0124] MEM6

[0125] An MEM6 sequence according to the invention includes a nucleicacid sequence encoding a polypeptide related to the glycogen-binding,phosphatase 1 protein family. The nucleotide sequence [SEQ ID NO: 11] ofthe novel nucleic acid (Internal Identification No. AC016485_A) encodinga glycogen-binding protein phosphatase 1-like protein is shown in FIG.26. An Open Reading Frame (ORF) was identified beginning with an atginitiation codon and ending with a tag termination codon. Putativeuntranslated regions upstream from the initiation codon and downstreamfrom the termination codon are shown by underlining, and the start andstop codons are shown in bold letters. The amino acid sequence [SEQ IDNO: 12] of the encoded protein is presented using the one-letter code inFIG. 27.

[0126] In a search of sequence databases, it was found, for example,that the nucleic acid sequence [SEQ ID NO: 11 ] has 763 of 903 bases(84%) identical to a rat mRNA for protein phosphatase 1 (GL-subunit)(GENBANK-ID:Y18208; see, FIG. 28). The amino acid sequence [SEQ ID NO:12] of the protein of the invention was found to have 255 of 284 aminoacid residues (89%) identical to, and 270 of 284 residues (92%) positivewith, the 284 amino acid residue hepatic glycogen-binding subunitprotein phosphatase-1 from rat (ACC: Q63759; see, FIG. 29).

[0127] A multiple sequence alignment is illustrated in FIG. 30, with theprotein of the invention being shown on Line 2, in a ClustalW analysiscomparing the protein of the invention with related protein sequences.

[0128] The novel nucleic acid of the invention encoding aglycogen-binding protein phosphatase 1 includes the nucleic acid whosesequence [SEQ ID NO:11] is provided in FIG. 26, or a fragment thereof.The invention also includes a mutant or variant nucleic acid any ofwhose bases may be changed from the corresponding base shown in FIG. 26,while still encoding a protein that maintains its glycogen-bindingprotein phosphatase 1-like activities and physiological functions, or afragment of such a nucleic acid. The invention further includes nucleicacids whose sequences are complementary to those just described,including nucleic acid fragments that are complementary to any of thenucleic acids just described. The invention additionally includesnucleic acids or nucleic acid fragments, or complements thereto, whosestructures include chemical modifications. Such modifications include,by way of non-limiting example, modified bases, and nucleic acids whosesugar phosphate backbones are modified or derivatized. Thesemodifications are carried out at least in part to enhance the chemicalstability of the modified nucleic acid, such that they may be used, forexample, as antisense binding nucleic acids in therapeutic applicationsin a subject. In the mutant or variant nucleic acids, and theircomplements, up to 20% or more of the bases may be so changed.

[0129] The novel protein of the invention includes the glycogen-bindingprotein phosphatase 1-like protein whose sequence [SEQ ID NO:12] isprovided in FIG. 27. The invention also includes a mutant or variantprotein any of whose residues may be changed from the correspondingresidue shown in FIG. 27, while still encoding a protein that maintainsits glycogen binding protein phosphatase 1-like activities andphysiological functions, or a functional fragment thereof. In the mutantor variant protein, up to 20% or more of the residues may be so changed.The invention further encompasses antibodies and antibody fragments,such as F_(ab) or (F_(ab))₂, that bind immunospecifically to any of theproteins of the invention.

[0130] MEM7

[0131] An MEM7 sequence according to the invention includes a nucleicacid sequence encoding a polypeptide related to retinol-binding proteinfamily. The nucleotide sequence [SEQ ID NO: 13] of the nucleic acid(Internal Identification No. AC01 8653_A) encoding a novel proteinresembling retinol-binding protein is shown in FIG. 31. An Open ReadingFrame (ORF) was identified beginning with an atg initiation codon andending with a tga termination codon. Putative untranslated regionsupstream from the initiation codon and downstream from the terminationcodon are shown by underlining, and the start and stop codons are shownin bold letters. The amino acid sequence [SEQ ID NO: 14] of the encodedprotein is presented using the one-letter code in FIG. 32.

[0132] In a search of sequence databases, it was found, e.g., that theMEM7 amino acid sequence [SEQ ID NO: 14] of the protein of the inventionhad 68 of 70 amino acid residues (97%) identical to, and 70 of 70residues (100%) positive with, the Human cytostatin Iprotein(patp:W27561; see, FIG. 33).

[0133] The novel nucleic acid of the invention encoding a proteinresembling retinol-binding protein includes the nucleic acid whosesequence [SEQ ID NO:13] is provided in FIG. 31, or a fragment thereof.The invention also includes a mutant or variant nucleic acid any ofwhose bases may be changed from the corresponding base shown in FIG. 31,while still encoding a protein that maintains its proteins resemblingretinol-binding activities and physiological functions, or a fragment ofsuch a nucleic acid. The invention further includes nucleic acids whosesequences are complementary to those just described, including nucleicacid fragments that are complementary to any of the nucleic acids justdescribed. The invention additionally includes nucleic acids or nucleicacid fragments, or complements thereto, whose structures includechemical modifications. Such modifications include, by way ofnon-limiting example, modified bases, and nucleic acids whose sugarphosphate backbones are modified or derivatized. These modifications arecarried out at least in part to enhance the chemical stability of themodified nucleic acid, such that they may be used, for example, asantisense binding nucleic acids in therapeutic applications in asubject. In the mutant or variant nucleic acids, and their complements,up to 20% or more of the bases may be so changed.

[0134] The novel protein of the invention includes the proteinsresembling retinol-binding protein whose sequence [SEQ ID NO: 14] isprovided in FIG. 32. The invention also includes a mutant or variantprotein any of whose residues may be changed from the correspondingresidue shown in FIG. 32, while still encoding a protein that maintainsits proteins resembling retinol-binding activities and physiologicalfunctions, or a functional fragment thereof. In the mutant or variantprotein, up to 20% or more of the residues may be so changed. Theinvention further encompasses antibodies and antibody fragments, such asFab or (Fab)2, that bind immunospecifically to any of the proteins ofthe invention.

[0135] MEM8

[0136] An MEM8 sequence according to the invention includes a nucleicacid sequence encoding a polypeptide related to retinol-binding proteinfamily. The nucleotide sequence [SEQ ID NO:15] of the novel nucleic acid(designated CuraGen Acc. No. AC018653A_da1) encoding a novel proteinresembling retinol-binding protein is shown in FIG. 34. An Open ReadingFrame (ORF) was identified beginning with an atg initiation codon andending with a tga termination codon. Putative untranslated regionsupstream from the initiation codon and downstream from the terminationcodon are shown by underlining, and the start and stop codons are shownin bold letters. The amino acid sequence [SEQ ID NO: 16] of the encodedprotein is presented using the one-letter code in FIG. 35.

[0137] In both a database analysis (see, FIG. 36), the amino acidsequence [SEQ ID NO:16] of the protein of the invention was found tohave 135 of 135 amino acid residues (100%) positive with, and 133 of 135residues (98%) identical to, the 135 amino acid residue Human cytostatinIII protein (patp:W30891).

[0138] A multiple sequence alignment is illustrated in FIG. 37, with theprotein of the invention being shown on Line 2, in a ClustalW analysiscomparing the protein of the invention with related protein sequences.

[0139] The novel nucleic acid of the invention encoding a proteinresembling retinol-binding protein includes the nucleic acid whosesequence [SEQ ID NO:15] is provided in FIG. 34, or a fragment thereof.The invention also includes a mutant or variant nucleic acid any ofwhose bases may be changed from the corresponding base shown in FIG. 34,while still encoding a protein that maintains its proteins resemblingretinol-binding activities and physiological functions, or a fragment ofsuch a nucleic acid. The invention further includes nucleic acids whosesequences are complementary to those just described, including nucleicacid fragments that are complementary to any of the nucleic acids justdescribed. The invention additionally includes nucleic acids or nucleicacid fragments, or complements thereto, whose structures includechemical modifications. Such modifications include, by way ofnon-limiting example, modified bases, and nucleic acids whose sugarphosphate backbones are modified or derivatized. These modifications arecarried out at least in part to enhance the chemical stability of themodified nucleic acid, such that they may be used, for example, asantisense binding nucleic acids in therapeutic applications in asubject. In the mutant or variant nucleic acids, and their complements,up to 20% or more of the bases may be so changed.

[0140] The novel protein of the invention includes the proteinsresembling retinol-binding protein whose sequence [SEQ ID NO:16] isprovided in FIG. 35. The invention also includes a mutant or variantprotein any of whose residues may be changed from the correspondingresidue shown in FIG. 35, while still encoding a protein that maintainsits proteins resembling retinol-binding activities and physiologicalfunctions, or a functional fragment thereof. In the mutant or variantprotein, up to 20% or more of the residues may be so changed. Theinvention further encompasses antibodies and antibody fragments, such asF_(ab) or (F_(ab))_(2,) that bind immunospecifically to any of theproteins of the invention.

[0141] MEMX Nucleic Acids

[0142] The nucleic acids of the invention include those that encode aMEMX polypeptide or protein. As used herein, the terms polypeptide andprotein are interchangeable.

[0143] In some embodiments, a MEMX nucleic acid encodes a mature MEMXpolypeptide. As used herein, a “mature” form of a polypeptide or proteindescribed herein relates to the product of a naturally occurringpolypeptide or precursor form or proprotein. The naturally occurringpolypeptide, precursor or proprotein includes, by way of non-limitingexample, the full length gene product, encoded by the correspondinggene. Alternatively, it may be defined as the polypeptide, precursor orproprotein encoded by an open reading frame described herein. Theproduct “mature” form arises, again by way of non-limiting example, as aresult of one or more naturally occurring processing steps that may takeplace within the cell in which the gene product arises. Examples of suchprocessing steps leading to a “mature” form of a polypeptide or proteininclude the cleavage of the amino-terminal methionine residue encoded bythe initiation codon of an open reading frame, or the proteolyticcleavage of a signal peptide or leader sequence. Thus a mature formarising from a precursor polypeptide or protein that has residues 1 toN, where residue 1 is the amino-terminal methionine, would have residues2 through N remaining after removal of the amino-terminal methionine.Alternatively, a mature form arising from a precursor polypeptide orprotein having residues 1 to N, in which an amino-terminal signalsequence from residue 1 to residue M is cleaved, would have the residuesfrom residue M+1 to residue N remaining. Further as used herein, a“mature” form of a polypeptide or protein may arise from a step ofpost-translational modification other than a proteolytic cleavage event.Such additional processes include, by way of non-limiting example,glycosylation, myristoylation or phosphorylation. In general, a maturepolypeptide or protein may result from the operation of only one ofthese processes, or a combination of any of them.

[0144] Among the MEMX nucleic acids is the nucleic acid whose sequenceis provided in SEQ ID NO:1, 3, 5, 7, 9, 11, 13, or 15, or a fragmentthereof. Additionally, the invention includes mutant or variant nucleicacids of SEQ ID NO:1, 3, 5, 7, 9, 11, 13, or 15, or a fragment thereof,any of whose bases may be changed from the corresponding bases shown inSEQ ID NO:1, 3, 5, 7, 9, 11, 13, or 15, while still encoding a proteinthat maintains at least one of its MEMX-like activities andphysiological functions (i.e., modulating angiogenesis, neuronaldevelopment). The invention further includes the complement of thenucleic acid sequence of SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, , or 15,including fragments, derivatives, analogs and homologs thereof. Theinvention additionally includes nucleic acids or nucleic acid fragments,or complements thereto, whose structures include chemical modifications.

[0145] One aspect of the invention pertains to isolated nucleic acidmolecules that encode MEMX proteins or biologically active portionsthereof. Also included are nucleic acid fragments sufficient for use ashybridization probes to identify MEMX-encoding nucleic acids (e.g., MEMXMRNA) and fragments for use as polymerase chain reaction (PCR) primersfor the amplification or mutation of MEMX nucleic acid molecules. Asused herein, the term “nucleic acid molecule” is intended to include DNAmolecules (e.g., cDNA or genomic DNA), RNA molecules (e.g., mRNA),analogs of the DNA or RNA generated using nucleotide analogs, andderivatives, fragments and homologs thereof. The nucleic acid moleculecan be single-stranded or double-stranded, but preferably isdouble-stranded DNA.

[0146] The term “probes” refer to nucleic acid sequences of variablelength, preferably between at least about 10 nucleotides (nt), 100 nt,or as many as about, e.g., 6,000 nt, depending on use. Probes are usedin the detection of identical, similar, or complementary nucleic acidsequences. Longer length probes are usually obtained from a natural orrecombinant source, are highly specific and much slower to hybridizethan oligomers. Probes may be single- or double-stranded and designed tohave specificity in PCR, membrane-based hybridization technologies, orELISA-like technologies.

[0147] An “isolated” nucleic acid molecule is one that is separated fromother nucleic acid molecules that are present in the natural source ofthe nucleic acid. Examples of isolated nucleic acid molecules include,but are not limited to, recombinant DNA molecules contained in a vector,recombinant DNA molecules maintained in a heterologous host cell,partially or substantially purified nucleic acid molecules, andsynthetic DNA or RNA molecules. Preferably, an “isolated” nucleic acidis free of sequences which naturally flank the nucleic acid (i.e.,sequences located at the 5′ and 3′ ends of the nucleic acid) in thegenomic DNA of the organism from which the nucleic acid is derived. Forexample, in various embodiments, the isolated MEMX nucleic acid moleculecan contain less than about 50 kb, 25 kb, 5 kb, 4 kb, 3 kb, 2 kb, 1 kb,0.5 kb or 0.1 kb of nucleotide sequences which naturally flank thenucleic acid molecule in genomic DNA of the cell from which the nucleicacid is derived. Moreover, an “isolated” nucleic acid molecule, such asa cDNA molecule, can be substantially free of other cellular material orculture medium when produced by recombinant techniques, or of chemicalprecursors or other chemicals when chemically synthesized.

[0148] A nucleic acid molecule of the present invention, e.g., a nucleicacid molecule having the nucleotide sequence of SEQ ID NO:1, 3, 5, 7, 9,11, 13, or 15, or a complement of any of this nucleotide sequence, canbe isolated using standard molecular biology techniques and the sequenceinformation provided herein. Using all or a portion of the nucleic acidsequence of SEQ ID NO:1, 3, 5, 7, 9, 11, 13, or 15, as a hybridizationprobe, MEMX nucleic acid sequences can be isolated using standardhybridization and cloning techniques (e.g., as described in Sambrook etal., eds., MOLECULAR CLONING: A LABORATORY MANUAL 2^(nd) Ed., ColdSpring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989; andAusubel, et al., eds., CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, JohnWiley & Sons, New York, N.Y., 1993.)

[0149] A nucleic acid of the invention can be amplified using cDNA, mRNAor alternatively, genomic DNA, as a template and appropriateoligonucleotide primers according to standard PCR amplificationtechniques. The nucleic acid so amplified can be cloned into anappropriate vector and characterized by DNA sequence analysis.Furthermore, oligonucleotides corresponding to MEMX nucleotide sequencescan be prepared by standard synthetic techniques, e.g., using anautomated DNA synthesizer.

[0150] As used herein, the term “oligonucleotide” refers to a series oflinked nucleotide residues, which oligonucleotide has a sufficientnumber of nucleotide bases to be used in a PCR reaction. A shortoligonucleotide sequence may be based on, or designed from, a genomic orcDNA sequence and is used to amplify, confirm, or reveal the presence ofan identical, similar or complementary DNA or RNA in a particular cellor tissue. Oligonucleotides comprise portions of a nucleic acid sequencehaving about 10 nt, 50 nt, or 100 nt in length, preferably about 15 ntto 30 nt in length. In one embodiment, an oligonucleotide comprising anucleic acid molecule less than 100 nt in length would further compriseat lease 6 contiguous nucleotides of SEQ ID NO:1, 3, 5, 7, 9, 11, 13, or15, or a complement thereof. Oligonucleotides may be chemicallysynthesized and may be used as probes.

[0151] In another embodiment, an isolated nucleic acid molecule of theinvention comprises a nucleic acid molecule that is a complement of thenucleotide sequence shown in SEQ ID NO:1, 3, 5, 7, 9, 11, 13, or 15, ora portion of this nucleotide sequence. A nucleic acid molecule that iscomplementary to the nucleotide sequence shown in SEQ ID NO:1, 3, 5, 7,9, 11, 13, or 15, is one that is sufficiently complementary to thenucleotide sequence shown in SEQ ID NO:1, 3, 5, 7, 9, 11, 13, or 15 thatit can hydrogen bond with little or no mismatches to the nucleotidesequence shown in SEQ ID NO:1, 3, 5, 7, 9, 11, 13, or 15, therebyforming a stable duplex.

[0152] As used herein, the term “complementary” refers to Watson-Crickor Hoogsteen base pairing between nucleotide units of a nucleic acidmolecule, and the term “binding” means the physical or chemicalinteraction between two polypeptides or compounds or associatedpolypeptides or compounds or combinations thereof. Binding includesionic, non-ionic, Von der Waals, hydrophobic interactions, etc. Aphysical interaction can be either direct or indirect. Indirectinteractions may be through or due to the effects of another polypeptideor compound. Direct binding refers to interactions that do not takeplace through, or due to, the effect of another polypeptide or compound,but instead are without other substantial chemical intermediates.

[0153] Moreover, the nucleic acid molecule of the invention can compriseonly a portion of the nucleic acid sequence of SEQ ID NO:1, 3, 5, 7, 9,11, 13, or 15, e.g., a fragment that can be used as a probe or primer,or a fragment encoding a biologically active portion of MEMX. Fragmentsprovided herein are defined as sequences of at least 6 (contiguous)nucleic acids or at least 4 (contiguous) amino acids, a lengthsufficient to allow for specific hybridization in the case of nucleicacids or for specific recognition of an epitope in the case of aminoacids, respectively, and are at most some portion less than a fulllength sequence. Fragments may be derived from any contiguous portion ofa nucleic acid or amino acid sequence of choice. Derivatives are nucleicacid sequences or amino acid sequences formed from the native compoundseither directly or by modification or partial substitution. Analogs arenucleic acid sequences or amino acid sequences that have a structuresimilar to, but not identical to, the native compound but differs fromit in respect to certain components or side chains. Analogs may besynthetic or from a different evolutionary origin and may have a similaror opposite metabolic activity compared to wild-type.

[0154] Derivatives and analogs may be full length or other than fulllength, if the derivative or analog contains a modified nucleic acid oramino acid, as described below. Derivatives or analogs of the nucleicacids or proteins of the invention include, but are not limited to,molecules comprising regions that are substantially homologous to thenucleic acids or proteins of the invention, in various embodiments, byat least about 70%, 80%, 82%, 90%, 92%, 98%, or even 99% identity (witha preferred identity of 80-99%) over a nucleic acid or amino acidsequence of identical size or when compared to an aligned sequence inwhich the alignment is done by a computer homology program known in theart, or whose encoding nucleic acid is capable of hybridizing to thecomplement of a sequence encoding the aforementioned proteins understringent, moderately stringent, or low stringent conditions. See e.g.Ausubel, et al., CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley &Sons, New York, N.Y., 1993, and below. An exemplary program is the Gapprogram (Wisconsin Sequence Analysis Package, Version 8 for UNIX,Genetics Computer Group, University Research Park, Madison, Wis.) usingthe default settings, which uses the algorithm of Smith and Waterman(1981. Adv. Appl. Math.2: 482-489, which is incorporated herein byreference in its entirety).

[0155] A “homologous nucleic acid sequence” or “homologous amino acidsequence,” or variations thereof, refer to sequences characterized by ahomology at the nucleotide level or amino acid level as discussed above.Homologous nucleotide sequences encode those sequences coding forisoforms of a MEMX polypeptide. Isoforms can be expressed in differenttissues of the same organism as a result of, for example, alternativesplicing of RNA. Alternatively, isoforms can be encoded by differentgenes. In the present invention, homologous nucleotide sequences includenucleotide sequences encoding for a MEMX polypeptide of species otherthan humans, including, but not limited to, mammals, and thus caninclude, e.g., mouse, rat, rabbit, dog, cat cow, horse, and otherorganisms. Homologous nucleotide sequences also include, but are notlimited to, naturally occurring allelic variations and mutations of thenucleotide sequences set forth herein. A homologous nucleotide sequencedoes not, however, include the nucleotide sequence encoding human MEMXprotein. Homologous nucleic acid sequences include those nucleic acidsequences that encode conservative amino acid substitutions (see below)in SEQ ID NO:2, 4, 6, 8, 10, 12, 14, or 16, as well as a polypeptidehaving MEMX activity. Biological activities of the MEMX proteins aredescribed below. A homologous amino acid sequence does not encode theamino acid sequence of a human MEMX polypeptide.

[0156] The nucleotide sequence determined from the cloning of the humanMEMX gene allows for the generation of probes and primers designed foruse in identifying and/or cloning MEMX homologues in other cell types,e.g., from other tissues, as well as MEMX homologues from other mammals.The probe/primer typically comprises a substantially purifiedoligonucleotide. The oligonucleotide typically comprises a region ofnucleotide sequence that hybridizes under stringent conditions to atleast about 12, 25, 50, 100, 150, 200, 250, 300, 350 or 400 or moreconsecutive sense strand nucleotide sequence of SEQ ID NO:1, 3, 5, 7, 9,11, 13, or 15; or an anti-sense strand nucleotide sequence of SEQ IDNO:1, 3, 5, 7, 9, 11, 13, or 15, or of a naturally occurring mutant ofSEQ ID NO:1, 3, 5, 7, 9, 11, 13, or 15.

[0157] Probes based upon the human MEMX nucleotide sequence can be usedto detect transcripts or genomic sequences encoding the same orhomologous proteins. In various embodiments, the probe further comprisesa label group attached thereto, e.g., the label group can be aradioisotope, a fluorescent compound, an enzyme, or an enzyme co-factor.Such probes can be used as a part of a diagnostic test kit foridentifying cells or tissue which misexpress a MEMX protein, such as bymeasuring a level of a MEMX-encoding nucleic acid in a sample of cellsfrom a subject e.g., detecting MEMX mRNA levels or determining whether agenomic MEMX gene has been mutated or deleted.

[0158] A “polypeptide having a biologically active portion of MEMX”refers to polypeptides exhibiting activity similar, but not necessarilyidentical to, an activity of a polypeptide of the present invention,including mature forms, as measured in a particular biological assay,with or without dose dependency. A nucleic acid fragment encoding a“biologically active portion of MEMX” can be prepared by isolating aportion of SEQ ID NO:1, 3, 5, 7, 9, 11, 13, or 15, that encodes apolypeptide having a MEMX biological activity (biological activities ofthe MEMX proteins are described below), expressing the encoded portionof MEMX protein (e.g., by recombinant expression in vitro) and assessingthe activity of the encoded portion of MEMX. For example, a nucleic acidfragment encoding a biologically active portion of MEMX can optionallyinclude an ATP-binding domain. In another embodiment, a nucleic acidfragment encoding a biologically active portion of MEMX includes one ormore regions.

[0159] MEMX Variants

[0160] The invention further encompasses nucleic acid molecules thatdiffer from the nucleotide sequences shown in SEQ ID NO:1, 3, 5, 7, 9,11, 13, or 15 due to the degeneracy of the genetic code. These nucleicacids thus encode the same MEMX protein as that encoded by thenucleotide sequence shown in SEQ ID NO:1, 3, 5, 7, 9, 11, 13, or 15,e.g., the polypeptide of SEQ ID NO:2, 4, 6, 8, 10, 12, 14, or 16.

[0161] In addition to the human MEMX nucleotide sequence shown in SEQ IDNO:1, 3, 5, 7, 9, 11, 13, or 15, it will be appreciated by those skilledin the art that DNA sequence polymorphisms that lead to changes in theamino acid sequences of MEMX may exist within a population (e.g., thehuman population). Such genetic polymorphism in the MEMX gene may existamong individuals within a population due to natural allelic variation.As used herein, the terms “gene” and “recombinant gene” refer to nucleicacid molecules comprising an open reading frame encoding a MEMX protein,preferably a mammalian MEMX protein. Such natural allelic variations cantypically result in 1-20% variance in the nucleotide sequence of theMEMX gene. Any and all such nucleotide variations and resulting aminoacid polymorphisms in MEMX that are the result of natural allelicvariation and that do not alter the functional activity of MEMX areintended to be within the scope of the invention.

[0162] Moreover, nucleic acid molecules encoding MEMX proteins fromother species, and thus that have a nucleotide sequence that differsfrom the human sequence of SEQ ID NO:1, 3, 5, 7, 9, 11, 13, or 15 areintended to be within the scope of the invention. Nucleic acid moleculescorresponding to natural allelic variants and homologues of the MEMXcDNAs of the invention can be isolated based on their homology to thehuman MEMX nucleic acids disclosed herein using the human cDNAs, or aportion thereof, as a hybridization probe according to standardhybridization techniques under stringent hybridization conditions. Forexample, a soluble human MEMX cDNA can be isolated based on its homologyto human membrane-bound MEMX. Likewise, a membrane-bound human MEMX cDNAcan be isolated based on its homology to soluble human MEMX.

[0163] Accordingly, in another embodiment, an isolated nucleic acidmolecule of the invention is at least 6 nucleotides in length andhybridizes under stringent conditions to the nucleic acid moleculecomprising the nucleotide sequence of SEQ ID NO:1, 3, 5, 7, 9, 11, 13,or 15. In another embodiment, the nucleic acid is at least 10, 25, 50,100, 250, 500 or 750 nucleotides in length. In another embodiment, anisolated nucleic acid molecule of the invention hybridizes to the codingregion. As used herein, the term “hybridizes under stringent conditions”is intended to describe conditions for hybridization and washing underwhich nucleotide sequences at least 60% homologous to each othertypically remain hybridized to each other.

[0164] Homologs (i.e., nucleic acids encoding MEMX proteins derived fromspecies other than human) or other related sequences (e.g., paralogs)can be obtained by low, moderate or high stringency hybridization withall or a portion of the particular human sequence as a probe usingmethods well known in the art for nucleic acid hybridization andcloning.

[0165] As used herein, the phrase “stringent hybridization conditions”refers to conditions under which a probe, primer or oligonucleotide willhybridize to its target sequence, but to no other sequences. Stringentconditions are sequence-dependent and will be different in differentcircumstances. Longer sequences hybridize specifically at highertemperatures than shorter sequences. Generally, stringent conditions areselected to be about 5° C. lower than the thermal melting point (Tm) forthe specific sequence at a defined ionic strength and pH. The Tm is thetemperature (under defined ionic strength, pH and nucleic acidconcentration) at which 50% of the probes complementary to the targetsequence hybridize to the target sequence at equilibrium. Since thetarget sequences are generally present at excess, at Tm, 50% of theprobes are occupied at equilibrium. Typically, stringent conditions willbe those in which the salt concentration is less than about 1.0 M sodiumion, typically about 0.01 to 1.0 M sodium ion (or other salts) at pH 7.0to 8.3 and the temperature is at least about 30° C. for short probes,primers or oligonucleotides (e.g., 10 nt to 50 nt) and at least about60° C. for longer probes, primers and oligonucleotides. Stringentconditions may also be achieved with the addition of destabilizingagents, such as formamide.

[0166] Stringent conditions are known to those skilled in the art andcan be found in CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley &Sons, N.Y. (1989), 6.3.1-6.3.6. Preferably, the conditions are such thatsequences at least about 620%, 70%, 72%, 82%, 90%, 92%, 98%, or 99%homologous to each other typically remain hybridized to each other. Anon-limiting example of stringent hybridization conditions ishybridization in a high salt buffer comprising 6×SSC, 50 mM Tris-HCl (pH7.5), 1 mM EDTA, 0.02% PVP, 0.02% Ficoll, 0.02% BSA, and 500 mg/mldenatured salmon sperm DNA at 65° C. This hybridization is followed byone or more washes in 0.2×SSC, 0.01% BSA at 50° C. An isolated nucleicacid molecule of the invention that hybridizes under stringentconditions to the sequence of SEQ ID NO:1, 3, 5, 7, 9, 11, 13, or 15,corresponds to a naturally occurring nucleic acid molecule. As usedherein, a “naturally-occurring” nucleic acid molecule refers to an RNAor DNA molecule having a nucleotide sequence that occurs in nature(e.g., encodes a natural protein).

[0167] In a second embodiment, a nucleic acid sequence that ishybridizable to the nucleic acid molecule comprising the nucleotidesequence of SEQ ID NO:1, 3, 5, 7, 9, 11, 13, or 15, or fragments,analogs or derivatives thereof, under conditions of moderate stringencyis provided. A non-limiting example of moderate stringency hybridizationconditions are hybridization in 6×SSC, 5×Denhardt's solution, 0.20% SDSand 100 mg/ml denatured salmon sperm DNA at 55° C., followed by one ormore washes in 1×SSC, 0.1% SDS at 37° C. Other conditions of moderatestringency that may be used are well known in the art. See, e.g.,Ausubel et al. (eds.), 1993, CURRENT PROTOCOLS IN MOLECULAR BIOLOGY,John Wiley & Sons, NY, and Kriegler, 1990, GENE TRANSFER AND ExPRESSION,A LABORATORY MANUAL, Stockton Press, NY.

[0168] In a third embodiment, a nucleic acid that is hybridizable to thenucleic acid molecule comprising the nucleotide sequence of SEQ ID NO:1,3, 5, 7, 9, 11, 13, or 15, or fragments, analogs or derivatives thereof,under conditions of low stringency, is provided. A non-limiting exampleof low stringency hybridization conditions are hybridization in 320%formamide, 5X SSC, 50 mM Tris-HCl (pH 7.5), 5 mM EDTA, 0.02% PVP, 0.02%Ficoll, 0.2% BSA, 100 mg/ml denatured salmon sperm DNA, 10% (wt/vol)dextran sulfate at 40° C., followed by one or more washes in 2×SSC, 25mM Tris-HCl (pH 7.4), 5 mM EDTA, and 0.1% SDS at 50° C. Other conditionsof low stringency that may be used are well known in the art (e.g., asemployed for cross-species hybridizations). See, e.g., Ausubel et al.(eds.), 1993, CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons,NY, and Kriegler, 1990, GENE TRANSFER AND EXPRESSION, A LABORATORYMANUAL, Stockton Press, NY; Shilo and Weinberg, 1981, Proc Natl Acad SciUSA 78: 6789-6792.

[0169] I. Conservative Mutations

[0170] In addition to naturally-occurring allelic variants of the MEMXsequence that may exist in the population, the skilled artisan willfurther appreciate that changes can be introduced by mutation into thenucleotide sequence of SEQ ID NO:1, 3, 5, 7, 9, 11, 13, or 15, therebyleading to changes in the amino acid sequence of the encoded MEMXprotein, without altering the functional ability of the MEMX protein.For example, nucleotide substitutions leading to amino acidsubstitutions at “non-essential” amino acid residues can be made in thesequence of SEQ ID NO:1, 3, 5, 7, 9, 11, 13, or 15. A “non-essential”amino acid residue is a residue that can be altered from the wild-typesequence of MEMX without altering the biological activity, whereas an“essential” amino acid residue is required for biological activity. Forexample, amino acid residues that are conserved among the MEMX proteinsof the present invention, are predicted to be particularly unamenable toalteration.

[0171] Another aspect of the invention pertains to nucleic acidmolecules encoding MEMX proteins that contain changes in amino acidresidues that are not essential for activity. Such MEMX proteins differin amino acid sequence from SEQ ID NO:2, 4, 6, 8, 10, 12, 14, or 16, yetretain biological activity. In one embodiment, the isolated nucleic acidmolecule comprises a nucleotide sequence encoding a protein, wherein theprotein comprises an amino acid sequence at least about 720% homologousto the amino acid sequence of SEQ ID NO:2, 4, 6, 8, 10, 12, 14, or 16.Preferably, the protein encoded by the nucleic acid is at least about80% homologous to SEQ ID NO:2, 4, 6, 8, 10, 12, 14, or 16, morepreferably at least about 90%, 92%, 98%, and most preferably at leastabout 99% homologous to SEQ ID NO:2, 4, 6, 8, 10, 12, 14, or 16.

[0172] An isolated nucleic acid molecule encoding a MEMX proteinhomologous to the protein of can be created by introducing one or morenucleotide substitutions, additions or deletions into the nucleotidesequence of SEQ ID NO:1, 3, 5, 7, 9, 11, 13, or 15, such that one ormore amino acid substitutions, additions or deletions are introducedinto the encoded protein.

[0173] Mutations can be introduced into the nucleotide sequence of SEQID NO:1, 3, 5, 7, 9, 11, 13, or 15 by standard techniques, such assite-directed mutagenesis and PCR-mediated mutagenesis. Preferably,conservative amino acid substitutions are made at one or more predictednon-essential amino acid residues. A “conservative amino acidsubstitution” is one in which the amino acid residue is replaced with anamino acid residue having a similar side chain. Families of amino acidresidues having similar side chains have been defined in the art. Thesefamilies include amino acids with basic side chains (e.g., lysine,arginine, histidine), acidic side chains (e.g., aspartic acid, glutamicacid), uncharged polar side chains (e.g., glycine, asparagine,glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains(e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine,methionine, tryptophan), beta-branched side chains (e.g., threonine,valine, isoleucine) and aromatic side chains (e.g., tyrosine,phenylalanine, tryptophan, histidine). Thus, a predicted nonessentialamino acid residue in MEMX is replaced with another amino acid residuefrom the same side chain family. Alternatively, in another embodiment,mutations can be introduced randomly along all or part of a MEMX codingsequence, such as by saturation mutagenesis, and the resultant mutantscan be screened for MEMX biological activity to identify mutants thatretain activity. Following mutagenesis of SEQ ID NO:1, 3, 5, 7, 9, 11,13, or 15, the encoded protein can be expressed by any recombinanttechnology known in the art and the activity of the protein can bedetermined.

[0174] In one embodiment, a mutant MEMX protein can be assayed for: (i)the ability to form protein:protein interactions with other MEMXproteins, other cell-surface proteins, or biologically active portionsthereof; (ii) complex formation between a mutant MEMX protein and a MEMXreceptor; (iii) the ability of a mutant MEMX protein to bind to anintracellular target protein or biologically active portion thereof;(e.g., avidin proteins); (iv) the ability to bind MEMX protein; or (v)the ability to specifically bind an anti-MEMX protein antibody.

[0175] Antisense MEMX Nucleic Acids

[0176] Another aspect of the invention pertains to isolated antisensenucleic acid molecules that are hybridizable to or complementary to thenucleic acid molecule comprising the nucleotide sequence of SEQ ID NO:1,3, 5, 7, 9, 11, 13, or 15, or fragments, analogs or derivatives thereof.An “antisense” nucleic acid comprises a nucleotide sequence that iscomplementary to a “sense” nucleic acid encoding a protein, e.g.,complementary to the coding strand of a double-stranded cDNA molecule orcomplementary to an mRNA sequence. In specific aspects, antisensenucleic acid molecules are provided that comprise a sequencecomplementary to at least about 10, 25, 50, 100, 250 or 500 nucleotidesor an entire MEMX coding strand, or to only a portion thereof. Nucleicacid molecules encoding fragments, homologs, derivatives and analogs ofa MEMX protein of SEQ ID NO:2, 4, 6, 8, 10, 12, 14, or 16, or antisensenucleic acids complementary to a MEMX nucleic acid sequence of SEQ IDNO:1, 3, 5, 7, 9, 11, 13, or 15 are additionally provided.

[0177] In one embodiment, an antisense nucleic acid molecule isantisense to a “coding region” of the coding strand of a nucleotidesequence encoding MEMX. The term “coding region” refers to the region ofthe nucleotide sequence comprising codons which are translated intoamino acid residues (e.g, the protein coding region of human MEMXcorresponds to SEQ ID NO:2, 4, 6, 8, 10, 12, 14, or 16). In anotherembodiment, the antisense nucleic acid molecule is antisense to a“non-coding region” of the coding strand of a nucleotide sequenceencoding MEMX. The term “non-coding region” refers to 5′ and 3′sequences which flank the coding region that are not translated intoamino acids (i.e., also referred to as 5′ and 3′ untranslated regions).

[0178] Given the coding strand sequences encoding MEMX disclosed herein(e.g., SEQ ID NO:1, 3, 5, 7, 9, 11, 13, or 15), antisense nucleic acidsof the invention can be designed according to the rules of Watson andCrick or Hoogsteen base pairing. The antisense nucleic acid molecule canbe complementary to the entire coding region of MEMX mRNA, but morepreferably is an oligonucleotide that is antisense to only a portion ofthe coding or non-coding region of MEMX mRNA. For example, the antisenseoligonucleotide can be complementary to the region surrounding thetranslation start site of MEMX mRNA. An antisense oligonucleotide canbe, for example, about 5, 10, 15, 20, 25, 30, 35, 40, 45 or 50nucleotides in length. An antisense nucleic acid of the invention can beconstructed using chemical synthesis or enzymatic ligation reactionsusing procedures known in the art. For example, an antisense nucleicacid (e.g., an antisense oligonucleotide) can be chemically synthesizedusing naturally occurring nucleotides or variously modified nucleotidesdesigned to increase the biological stability of the molecules or toincrease the physical stability of the duplex formed between theantisense and sense nucleic acids, e.g., phosphorothioate derivativesand acridine substituted nucleotides can be used.

[0179] Examples of modified nucleotides that can be used to generate theantisense nucleic acid include: 5-fluorouracil, 5-bromouracil,5-chlorouracil, 5-iodouracil, hypoxanthine, xanthine, 4-acetylcytosine,5-(carboxyhydroxylmethyl) uracil,5-carboxymethylaminomethyl-2-thiouridine,5-carboxymethylaminomethyluracil, dihydrouracil,beta-D-galactosylqueosine, inosine, N6-isopentenyladenine,1-methylguanine, 1-methylinosine, 2,2-dimethylguanine, 2-methyladenine,2-methylguanine, 3-methylcytosine, 5-methylcytosine, N6-adenine,7-methylguanine, 5-methylaminomethyluracil,5-methoxyaminomethyl-2-thiouracil, beta-D-mannosylqueosine,5′-methoxycarboxymethyluracil, 5-methoxyuracil,2-methylthio-N6-isopentenyladenine, uracil-5-oxyacetic acid (v),wybutoxosine, pseudouracil, queosine, 2-thiocytosine,5-methyl-2-thiouracil, 2-thiouracil, 4-thiouracil, 5-methyluracil,uracil-5-oxyacetic acid methylester, uracil-5-oxyacetic acid (v),5-methyl-2-thiouracil, 3-(3-amino-3-N-2-carboxypropyl) uracil, (acp3)w,and 2,6-diaminopurine. Alternatively, the antisense nucleic acid can beproduced biologically using an expression vector into which a nucleicacid has been subcloned in an antisense orientation (i.e., RNAtranscribed from the inserted nucleic acid will be of an antisenseorientation to a target nucleic acid of interest, described further inthe following subsection).

[0180] The antisense nucleic acid molecules of the invention aretypically administered to a subject or generated in situ such that theyhybridize with or bind to cellular mRNA and/or genomic DNA encoding aMEMX protein to thereby inhibit expression of the protein, e.g., byinhibiting transcription and/or translation. The hybridization can be byconventional nucleotide complementarity to form a stable duplex, or, forexample, in the case of an antisense nucleic acid molecule that binds toDNA duplexes, through specific interactions in the major groove of thedouble helix. An example of a route of administration of antisensenucleic acid molecules of the invention includes direct injection at atissue site. Alternatively, antisense nucleic acid molecules can bemodified to target selected cells and then administered systemically.For example, for systemic administration, antisense molecules can bemodified such that they specifically bind to receptors or antigensexpressed on a selected cell surface, e.g., by linking the antisensenucleic acid molecules to peptides or antibodies that bind to cellsurface receptors or antigens. The antisense nucleic acid molecules canalso be delivered to cells using the vectors described herein. Toachieve sufficient intracellular concentrations of antisense molecules,vector constructs in which the antisense nucleic acid molecule is placedunder the control of a strong pol II or pol III promoter are preferred.

[0181] In yet another embodiment, the antisense nucleic acid molecule ofthe invention is an α-anomeric nucleic acid molecule. An α-anomericnucleic acid molecule forms specific double-stranded hybrids withcomplementary RNA in which, contrary to the usual β-units, the strandsrun parallel to each other (see, Gaultier, et al. 1987. Nucl. Acids Res.15: 6625-6641). The antisense nucleic acid molecule can also comprise a2′-o-methylribonucleotide (see, Inoue, et al., 1987. Nucl. Acids Res.15: 6131-6148) or a chimeric RNA -DNA analogue (see, Inoue, et al.,.1987. FEBS Lett. 215: 327-330).

[0182] Such modifications include, by way of non-limiting example,modified bases, and nucleic acids whose sugar phosphate backbones aremodified or derivatized. These modifications are carried out at least inpart to enhance the chemical stability of the modified nucleic acid,such that they may be used, for example, as antisense binding nucleicacids in therapeutic applications in a subject.

[0183] MEMX Ribozymes and PNA Moieties

[0184] In still another embodiment, an antisense nucleic acid of theinvention is a ribozyme. Ribozymes are catalytic RNA molecules withribonuclease activity that are capable of cleaving a single-strandednucleic acid, such as a mRNA, to which they have a complementary region.Thus, ribozymes (e.g., hammerhead ribozymes; see, Haselhoff and Gerlach,1988. Nature 334: 585-591) can be used to catalytically-cleave MEMX mRNAtranscripts to thereby inhibit translation of MEMX mRNA. A ribozymehaving specificity for a MEMX-encoding nucleic acid can be designedbased upon the nucleotide sequence of a MEMX DNA disclosed herein (i.e.,SEQ ID NO:1, 3, 5, 7, 9, 11, 13, or 15). For example, a derivative of aTetrahymena L-19 IVS RNA can be constructed in which the nucleotidesequence of the active site is complementary to the nucleotide sequenceto be cleaved in a MEMX-encoding mRNA. See, e.g, Cech, et al. U.S. Pat.No. 4,987,071; and Cech, et al., U.S. Pat. No. 5,116,742. Alternatively,MEMX mRNA can be used to select a catalytic RNA having a specificribonuclease activity from a pool of RNA molecules. See, e.g., Bartel,et al., 1993. Science 261: 1411-1418.

[0185] Alternatively, MEMX gene expression can be inhibited by targetingnucleotide sequences complementary to the regulatory region of the MEMX(e.g., the MEMX promoter and/or enhancers) to form triple helicalstructures that prevent transcription of the MEMX gene in target cells.See, generally, Helene, 1991. Anticancer Drug Des. 6: 569-584; Helene.et al. 1992. Ann. N.Y. Acad. Sci. 660: 27-36; and Maher. 1992. Bioassays14: 807-815.

[0186] In various embodiments, the nucleic acids of MEMX can be modifiedat the base moiety, sugar moiety or phosphate backbone to improve, e.g.,the stability, hybridization, or solubility of the molecule. Forexample, the deoxyribose phosphate backbone of the nucleic acids can bemodified to generate peptide nucleic acids (see, Hyrup, et al. 1996.Bioorg Med. Chem. 4: 5-23). As used herein, the terms “peptide nucleicacids” or “PNAs” refer to nucleic acid mimics, e.g., DNA mimics, inwhich the deoxyribose phosphate backbone is replaced by a pseudopeptidebackbone and only the four natural nucleobases are retained. The neutralbackbone of PNAs has been shown to allow for specific hybridization toDNA and RNA under conditions of low ionic strength. The synthesis of PNAoligomers can be performed using standard solid phase peptide synthesisprotocols as described in Hyrup, et al., 1996. supra; Perry-O'Keefe, etal., 1996. Proc. Natl. Acad. Sci. USA 93: 14670-14675.

[0187] PNAs of MEMX can be used in therapeutic and diagnosticapplications. For example, PNAs can be used as antisense or antigeneagents for sequence-specific modulation of gene expression by, e.g.,inducing transcription or translation arrest or inhibiting replication.PNAs of MEMX can also be used, e.g., in the analysis of single base pairmutations in a gene by, e.g., PNA directed PCR clamping; as artificialrestriction enzymes when used in combination with other enzymes, e.g.,S1 nucleases (Hyrup, 1996. supra); or as probes or primers for DNAsequence and hybridization (Hyrup, 1996 and Perry-O'Keefe, 1996.,supra).

[0188] In another embodiment, PNAs of MEMX can be modified, e.g., toenhance their stability or cellular uptake, by attaching lipophilic orother helper groups to PNA, by the formation of PNA-DNA chimeras, or bythe use of liposomes or other techniques of drug delivery known in theart. For example, PNA-DNA chimeras of MEMX can be generated that maycombine the advantageous properties of PNA and DNA. Such chimeras allowDNA recognition enzymes, e.g., RNase H and DNA polymerases, to interactwith the DNA portion while the PNA portion would provide high bindingaffinity and specificity. PNA-DNA chimeras can be linked using linkersof appropriate lengths selected in terms of base stacking, number ofbonds between the nucleobases, and orientation. The synthesis of PNA-DNAchimeras can be performed (see, e.g., Finn, et al., 1996. Nucl. AcidsRes. 24: 3357-3363. For example, a DNA chain can be synthesized on asolid support using standard phosphoramidite coupling chemistry, andmodified nucleoside analogs, e.g., 5′-(4-methoxytrityl)amino-5′-deoxy-thymidine phosphoramidite, can be used between the PNAand the 5′ end of DNA (see, Mag, et al., 1989. Nucl. Acids Res. 17:5973-5988). PNA monomers are then coupled in a stepwise manner toproduce a chimeric molecule with a 5′ PNA segment and a 3′ DNA segment(Finn, et al., 1996., supra). Alternatively, chimeric molecules can besynthesized with a 5′ DNA segment and a 3′ PNA segment (see, Petersen,et al., 1975. Bioorg. Med. Chem. Lett. 5: 1119-1124.

[0189] In other embodiments, the oligonucleotide may include otherappended groups such as peptides (e.g., for targeting host cellreceptors in vivo), or agents facilitating transport across the cellmembrane (see, e.g., Letsinger, et al., 1989. Proc. Natl. Acad. SciU.S.A. 86: 6553-6556; Lemaitre, et al., 1987. Proc. Natl. Acad. Sci. 84:648-652; PCT Publication No. WO88/09810) or the blood-brain barrier(see, e.g., PCT Publication No. WO 89/10134). In addition,oligonucleotides can be modified with hybridization triggered cleavageagents (see, e.g., Krol, et al., 1988. BioTechniques 6:958-976) orintercalating agents (see, e.g., Zon, 1988. Pharm. Res. 5: 539-549). Tothis end, the oligonucleotide may be conjugated to another molecule,e.g., a peptide, a hybridization triggered cross-linking agent, atransport agent, a hybridization-triggered cleavage agent, and the like.

[0190] MEMX Polypeptides

[0191] A MEMX polypeptide of the invention includes the MEMX-likeprotein whose sequence is provided in SEQ ID NO:2, 4, 6, 8, 10, 12, 14,or 16. The invention also includes a mutant or variant protein any ofwhose residues may be changed from the corresponding residue shown inSEQ ID NO:2, 4, 6, 8, 10, 12, 14, or 16, while still encoding a proteinthat maintains its MEMX-like activities and physiological functions, ora functional fragment thereof. In some embodiments, up to 20% or more ofthe residues may be so changed in the mutant or variant protein. In someembodiments, the MEMX polypeptide according to the invention is a maturepolypeptide.

[0192] In general, a MEMX -like variant that preserves MEMX-likefunction includes any variant in which residues at a particular positionin the sequence have been substituted by other amino acids, and furtherinclude the possibility of inserting an additional residue or residuesbetween two residues of the parent protein as well as the possibility ofdeleting one or more residues from the parent sequence. Any amino acidsubstitution, insertion, or deletion is encompassed by the invention. Infavorable circumstances, the substitution is a conservative substitutionas defined above.

[0193] One aspect of the invention pertains to isolated MEMX proteins,and biologically active portions thereof, or derivatives, fragments,analogs or homologs thereof. Also provided are polypeptide fragmentssuitable for use as immunogens to raise anti-MEMX antibodies. In oneembodiment, native MEMX proteins can be isolated from cells or tissuesources by an appropriate purification scheme using standard proteinpurification techniques. In another embodiment, MEMX proteins areproduced by recombinant DNA techniques. Alternative to recombinantexpression, a MEMX protein or polypeptide can be synthesized chemicallyusing standard peptide synthesis techniques.

[0194] An “isolated” or “purified” protein or biologically activeportion thereof is substantially free of cellular material or othercontaminating proteins from the cell or tissue source from which theMEMX protein is derived, or substantially free from chemical precursorsor other chemicals when chemically synthesized. The language“substantially free of cellular material” includes preparations of MEMXprotein in which the protein is separated from cellular components ofthe cells from which it is isolated or recombinantly produced. In oneembodiment, the language “substantially free of cellular material”includes preparations of MEMX protein having less than about 30% (by dryweight) of non-MEMX protein (also referred to herein as a “contaminatingprotein”), more preferably less than about 20% of non-MEMX protein,still more preferably less than about 10% of non-MEMX protein, and mostpreferably less than about 20% non-MEMX protein. When the MEMX proteinor biologically active portion thereof is recombinantly produced, it isalso preferably substantially free of culture medium, i.e., culturemedium represents less than about 20%, more preferably less than about10%, and most preferably less than about 20% of the volume of theprotein preparation.

[0195] The language “substantially free of chemical precursors or otherchemicals” includes preparations of MEMX protein in which the protein isseparated from chemical precursors or other chemicals that are involvedin the synthesis of the protein. In one embodiment, the language“substantially free of chemical precursors or other chemicals” includespreparations of MEMX protein having less than about 30% (by dry weight)of chemical precursors or non-MEMX chemicals, more preferably less thanabout 20% chemical precursors or non-MEMX chemicals, still morepreferably less than about 10% chemical precursors or non-MEMXchemicals, and most preferably less than about 20% chemical precursorsor non-MEMX chemicals.

[0196] Biologically active portions of a MEMX protein include peptidescomprising amino acid sequences sufficiently homologous to or derivedfrom the amino acid sequence of the MEMX protein, e.g., the amino acidsequence shown in SEQ ID NO:2, 4, 6, 8, 10, 12, 14, or 16, that includefewer amino acids than the full length MEMX proteins, and exhibit atleast one activity of a MEMX protein. Typically, biologically activeportions comprise a domain or motif with at least one activity of theMEMX protein. A biologically active portion of a MEMX protein can be apolypeptide which is, for example, 10, 25, 50, 100 or more amino acidsin length.

[0197] A biologically active portion of a MEMX protein of the presentinvention may contain at least one of the above-identified domainsconserved between the MEMX proteins, e.g. TSR modules. Moreover, otherbiologically active portions, in which other regions of the protein aredeleted, can be prepared by recombinant techniques and evaluated for oneor more of the functional activities of a native MEMX protein.

[0198] In an embodiment, the MEMX protein has an amino acid sequenceshown in SEQ ID NO:2, 4, 6, 8, 10, 12, 14, or 16. In other embodiments,the MEMX protein is substantially homologous to SEQ ID NO:2, 4, 6, 8,10, 12, 14, or 16 and retains the functional activity of the protein ofSEQ ID NO:2, 4, 6, 8, 10, 12, 14, or 16, yet differs in amino acidsequence due to natural allelic variation or mutagenesis, as describedin detail below. Accordingly, in another embodiment, the MEMX protein isa protein that comprises an amino acid sequence at least about 45%homologous, and more preferably about 55, 65, 70, 75, 80, 85, 90, 95,98, or even 99% homologous to the amino acid sequence of SEQ ID NO:2, 4,6, 8, 10, 12, 14, or 16 and retains the functional activity of the MEMXproteins of SEQ ID NO:2, 4, 6, 8, 10, 12, 14, or 16.

[0199] I. Determining Homology Between Two or More Amino Acid Sequences

[0200] To determine the percent homology of two amino acid sequences orof two nucleic acids, the sequences are aligned for optimal comparisonpurposes (e.g., gaps can be introduced in either of the sequences beingcompared for optimal alignment between the sequences). The amino acidresidues or nucleotides at corresponding amino acid positions ornucleotide positions are then compared. When a position in the firstsequence is occupied by the same amino acid residue or nucleotide as thecorresponding position in the second sequence, then the molecules arehomologous at that position (i.e., as used herein amino acid or nucleicacid “homology” is equivalent to amino acid or nucleic acid “identity”).

[0201] The nucleic acid sequence homology may be determined as thedegree of identity between two sequences. The homology may be determinedusing computer programs known in the art, such as GAP software providedin the GCG program package. See, Needleman and Wunsch 1970 J Mol Biol48: 443-453. Using GCG GAP software with the following settings fornucleic acid sequence comparison: GAP creation penalty of 5.0 and GAPextension penalty of 0.3, the coding region of the analogous nucleicacid sequences referred to above exhibits a degree of identitypreferably of at least 70%, 72%, 80%, 82%, 90%, 92%, 98%, or 99%, withthe CDS (encoding) part of the DNA sequence shown in SEQ ID NO:1, 3, 5,7, 9, 11, 13, or 15.

[0202] The term “sequence identity” refers to the degree to which twopolynucleotide or polypeptide sequences are identical on aresidue-by-residue basis over a particular region of comparison. Theterm “percentage of sequence identity” is calculated by comparing twooptimally aligned sequences over that region of comparison, determiningthe number of positions at which the identical nucleic acid base (e.g.,A, T, C, G, U, or I, in the case of nucleic acids) occurs in bothsequences to yield the number of matched positions, dividing the numberof matched positions by the total number of positions in the region ofcomparison (i.e., the window size), and multiplying the result by 100 toyield the percentage of sequence identity. The term “substantialidentity” as used herein denotes a characteristic of a polynucleotidesequence, wherein the polynucleotide comprises a sequence that has atleast 80 percent sequence identity, preferably at least 85 percentidentity and often 90 to 95 percent sequence identity, more usually atleast 99 percent sequence identity as compared to a reference sequenceover a comparison region. The term “percentage of positive residues” iscalculated by comparing two optimally aligned sequences over that regionof comparison, determining the number of positions at which theidentical and conservative amino acid substitutions, as defined above,occur in both sequences to yield the number of matched positions,dividing the number of matched positions by the total number ofpositions in the region of comparison (i.e., the window size), andmultiplying the result by 100 to yield the percentage of positiveresidues.

[0203] Chimeric and Fusion Proteins

[0204] The invention also provides MEMX chimeric or fusion proteins. Asused herein, a MEMX “chimeric protein” or “fusion protein” comprises aMEMX polypeptide operatively linked to a non-MEMX polypeptide. An “MEMXpolypeptide” refers to a polypeptide having an amino acid sequencecorresponding to MEMX, whereas a “non-MEMX polypeptide” refers to apolypeptide having an amino acid sequence corresponding to a proteinthat is not substantially homologous to the MEMX protein, e.g., aprotein that is different from the MEMX protein and that is derived fromthe same or a different organism. Within a MEMX fusion protein the MEMXpolypeptide can correspond to all or a portion of a MEMX protein. In oneembodiment, a MEMX fusion protein comprises at least one biologicallyactive portion of a MEMX protein. In another embodiment, a MEMX fusionprotein comprises at least two biologically active portions of a MEMXprotein. Within the fusion protein, the term “operatively linked” isintended to indicate that the MEMX polypeptide and the non-MEMXpolypeptide are fused in-frame to each other. The non-MEMX polypeptidecan be fused to the N-terminus or C-terminus of the MEMX polypeptide.

[0205] For example, in one embodiment a MEMX fusion protein comprises aMEMX polypeptide operably linked to the extracellular domain of a secondprotein. Such fusion proteins can be further utilized in screeningassays for compounds that modulate MEMX activity (such assays aredescribed in detail below).

[0206] In another embodiment, the fusion protein is a glutathioneS-transferase (GST)-MEMX fusion protein in which the MEMX sequences arefused to the carboxyl-terminus of the GST sequences. Such fusionproteins can facilitate the purification of recombinant MEMX.

[0207] In another embodiment, the fusion protein is aMEMX-immunoglobulin fusion protein in which the MEMX sequencescomprising one or more domains are fused to sequences derived from amember of the immunoglobulin protein family. The MEMX-immunoglobulinfusion proteins of the invention can be incorporated into pharmaceuticalcompositions and administered to a subject to inhibit an interactionbetween a MEMX ligand and a MEMX protein on the surface of a cell, tothereby suppress MEMX-mediated signal transduction in vivo. In onenon-limiting example, a contemplated MEMX ligand of the invention is theMEMX receptor. The MEMX-immunoglobulin fusion proteins can be used toaffect the bioavailability of a MEMX cognate ligand. Inhibition of theMEMX ligand/MEMX interaction may be useful therapeutically for both thetreatment of proliferative and differentiative disorders, e,g., canceras well as modulating (e.g., promoting or inhibiting) cell survival.Moreover, the MEMX-immunoglobulin fusion proteins of the invention canbe used as immunogens to produce anti-MEMX antibodies in a subject, topurify MEMX ligands, and in screening assays to identify molecules thatinhibit the interaction of MEMX with a MEMX ligand.

[0208] A MEMX chimeric or fusion protein of the invention can beproduced by standard recombinant DNA techniques. For example, DNAfragments coding for the different polypeptide sequences are ligatedtogether in-frame in accordance with conventional techniques, e.g., byemploying blunt-ended or stagger-ended termini for ligation, restrictionenzyme digestion to provide for appropriate termini, filling-in ofcohesive ends as appropriate, alkaline phosphatase treatment to avoidundesirable joining, and enzymatic ligation. In another embodiment, thefusion gene can be synthesized by conventional techniques includingautomated DNA synthesizers. Alternatively, PCR amplification of genefragments can be carried out using anchor primers that give rise tocomplementary overhangs between two consecutive gene fragments that cansubsequently be annealed and reamplified to generate a chimeric genesequence (see, for example, Ausubel, et al. (eds.) CURRENT PROTOCOLS INMOLECULAR BIOLOGY, John Wiley & Sons, 1992). Moreover, many expressionvectors are commercially available that already encode a fusion moiety(e.g., a GST polypeptide). A MEMX-encoding nucleic acid can be clonedinto such an expression vector such that the fusion moiety is linkedin-frame to the MEMX protein.

[0209] MEMX Agonists and Antagonists

[0210] The present invention also pertains to variants of the MEMXproteins that function as either MEMX agonists (i.e., mimetics) or asMEMX antagonists. Variants of the MEMX protein can be generated bymutagenesis, e.g., discrete point mutation or truncation of the MEMXprotein. An agonist of the MEMX protein can retain substantially thesame, or a subset of, the biological activities of the naturallyoccurring form of the MEMX protein. An antagonist of the MEMX proteincan inhibit one or more of the activities of the naturally occurringform of the MEMX protein by, for example, competitively binding to adownstream or upstream member of a cellular signaling cascade whichincludes the MEMX protein. Thus, specific biological effects can beelicited by treatment with a variant of limited function. In oneembodiment, treatment of a subject with a variant having a subset of thebiological activities of the naturally occurring form of the protein hasfewer side effects in a subject relative to treatment with the naturallyoccurring form of the MEMX proteins.

[0211] Variants of the MEMX protein that function as either MEMXagonists (mimetics) or as MEMX antagonists can be identified byscreening combinatorial libraries of mutants, e.g., truncation mutants,of the MEMX protein for MEMX protein agonist or antagonist activity. Inone embodiment, a variegated library of MEMX variants is generated bycombinatorial mutagenesis at the nucleic acid level and is encoded by avariegated gene library. A variegated library of MEMX variants can beproduced by, for example, enzymatically ligating a mixture of syntheticoligonucleotides into gene sequences such that a degenerate set ofpotential MEMX sequences is expressible as individual polypeptides, oralternatively, as a set of larger fusion proteins (e.g., for phagedisplay) containing the set of MEMX sequences therein. There are avariety of methods which can be used to produce libraries of potentialMEMX variants from a degenerate oligonucleotide sequence. Chemicalsynthesis of a degenerate gene sequence can be performed in an automaticDNA synthesizer, and the synthetic gene then ligated into an appropriateexpression vector. Use of a degenerate set of genes allows for theprovision, in one mixture, of all of the sequences encoding the desiredset of potential MEMX sequences. Methods for synthesizing degenerateoligonucleotides are known in the art (see, e.g., Narang 1983.Tetrahedron 39:3; Itakura, et al., 1984. Annual Rev. Biochem. 53: 323;Itakura, et al., 1984. Science 198:1056; Ike, et al., 1983. Nucl. AcidRes. 11:477.

[0212] I. Polypeptide Libraries

[0213] In addition, libraries of fragments of the MEMX protein codingsequences can be used to generate a variegated population of MEMXfragments for screening and subsequent selection of variants of an MEMXprotein. In one embodiment, a library of coding sequence fragments canbe generated by treating a double stranded PCR fragment of an MEMXcoding sequence with a nuclease under conditions wherein nicking occursonly about once per molecule, denaturing the double stranded DNA,renaturing the DNA to form double-stranded DNA that can includesense/antisense pairs from different nicked products, removing singlestranded portions from reformed duplexes by treatment with S₁ nuclease,and ligating the resulting fragment library into an expression vector.By this method, expression libraries can be derived which encodesN-terminal and internal fragments of various sizes of the MEMX proteins.

[0214] Various techniques are known in the art for screening geneproducts of combinatorial libraries made by point mutations ortruncation, and for screening cDNA libraries for gene products having aselected property. Such techniques are adaptable for rapid screening ofthe gene libraries generated by the combinatorial mutagenesis of MEMXproteins. The most widely used techniques, which are amenable to highthroughput analysis, for screening large gene libraries typicallyinclude cloning the gene library into replicable expression vectors,transforming appropriate cells with the resulting library of vectors,and expressing the combinatorial genes under conditions in whichdetection of a desired activity facilitates isolation of the vectorencoding the gene whose product was detected. Recursive ensemblemutagenesis (REM), a new technique that enhances the frequency offunctional mutants in the libraries, can be used in combination with thescreening assays to identify MEMX variants. See, e.g., Arkin andYourvan, 1992. Proc. Natl. Acad. Sci. USA 89: 7811-7815; Delgrave, etal., 1993. Protein Engineering 6:327-331.

[0215] Anti-MEMX Antibodies

[0216] Also included in the invention are antibodies to MEMX proteins,or fragments of MEMX proteins. The term “antibody” as used herein refersto immunoglobulin molecules and immunologically active portions ofimmunoglobulin (Ig) molecules, i.e., molecules that contain an antigenbinding site that specifically binds (immunoreacts with) an antigen.Such antibodies include, but are not limited to, polyclonal, monoclonal,chimeric, single chain, Fab, F_(ab′) and F_((ab′)2) fragments, and anF_(ab) expression library. In general, an antibody molecule obtainedfrom humans relates to any of the classes IgG, IgM, IgA, IgE and IgD,which differ from one another by the nature of the heavy chain presentin the molecule. Certain classes have subclasses as well, such as IgG₁,IgG₂, and others. Furthermore, in humans, the light chain may be a kappachain or a lambda chain. Reference herein to antibodies includes areference to all such classes, subclasses and types of human antibodyspecies.

[0217] An isolated MEMX-related protein of the invention may be intendedto serve as an antigen, or a portion or fragment thereof, andadditionally can be used as an immunogen to generate antibodies thatimmunospecifically bind the antigen, using standard techniques forpolyclonal and monoclonal antibody preparation. The full-length proteincan be used or, alternatively, the invention provides antigenic peptidefragments of the antigen for use as immunogens. An antigenic peptidefragment comprises at least 6 amino acid residues of the amino acidsequence of the full length protein, such as an amino acid sequenceshown in SEQ ID NO:2,4, 6, 8, 10, 12, 14, or 16, and encompasses anepitope thereof such that an antibody raised against the peptide forms aspecific immune complex with the full length protein or with anyfragment that contains the epitope. Preferably, the antigenic peptidecomprises at least 10 amino acid residues, or at least 15 amino acidresidues, or at least 20 amino acid residues, or at least 30 amino acidresidues. Preferred epitopes encompassed by the antigenic peptide areregions of the protein that are located on its surface; commonly theseare hydrophilic regions.

[0218] In certain embodiments of the invention, at least one epitopeencompassed by the antigenic peptide is a region of MEMX-related proteinthat is located on the surface of the protein, e.g., a hydrophilicregion. A hydrophobicity analysis of the human MEMX-related proteinsequence will indicate which regions of a MEMX-related protein areparticularly hydrophilic and, therefore, are likely to encode surfaceresidues useful for targeting antibody production. As a means fortargeting antibody production, hydropathy plots showing regions ofhydrophilicity and hydrophobicity may be generated by any method wellknown in the art, including, for example, the Kyte Doolittle or the HoppWoods methods, either with or without Fourier transformation. See, e.g,Hopp and Woods, 1981, Proc. Nat. Acad. Sci. USA 78: 3824-3828; Kyte andDoolittle, 1982. J. Mol. Biol. 157: 105-142, each of which isincorporated herein by reference in its entirety. Antibodies that arespecific for one or more domains within an antigenic protein, orderivatives, fragments, analogs or homologs thereof, are also providedherein.

[0219] A protein of the invention, or a derivative, fragment, analog,homolog or ortholog thereof, may be utilized as an immunogen in thegeneration of antibodies that immunospecifically bind these proteincomponents.

[0220] Various procedures known within the art may be used for theproduction of polyclonal or monoclonal antibodies directed against aprotein of the invention, or against derivatives, fragments, analogshomologs or orthologs thereof (see, for example, ANTIBODIES: ALABORATORY MANUAL, Harlow E, and Lane D, 1988, Cold Spring HarborLaboratory Press, Cold Spring Harbor, N.Y., incorporated herein byreference). Some of these antibodies are discussed below.

[0221] I. Polyclonal Antibodies

[0222] For the production of polyclonal antibodies, various suitablehost animals (e.g., rabbit, goat, mouse or other mammal) may beimmunized by one or more injections with the native protein, a syntheticvariant thereof, or a derivative of the foregoing. An appropriateimmunogenic preparation can contain, for example, the naturallyoccurring immunogenic protein, a chemically synthesized polypeptiderepresenting the immunogenic protein, or a recombinantly expressedimmunogenic protein. Furthermore, the protein may be conjugated to asecond protein known to be immunogenic in the mammal being immunized.Examples of such immunogenic proteins include but are not limited tokeyhole limpet hemocyanin, serum albumin, bovine thyroglobulin, andsoybean trypsin inhibitor. The preparation can further include anadjuvant. Various adjuvants used to increase the immunological responseinclude, but are not limited to, Freund's (complete and incomplete),mineral gels (e.g., aluminum hydroxide), surface active substances(e.g., lysolecithin, pluronic polyols, polyanions, peptides, oilemulsions, dinitrophenol, etc.), adjuvants usable in humans such asBacille Calmette-Guerin and Corynebacterium parvum, or similarimmunostimulatory agents. Additional examples of adjuvants which can beemployed include MPL-TDM adjuvant (monophosphoryl Lipid A, synthetictrehalose dicorynomycolate).

[0223] The polyclonal antibody molecules directed against theimmunogenic protein can be isolated from the mammal (e.g., from theblood) and further purified by well known techniques, such as affinitychromatography using protein A or protein G, which provide primarily theIgG fraction of immune serum. Subsequently, or alternatively, thespecific antigen which is the target of the immunoglobulin sought, or anepitope thereof, may be immobilized on a column to purify the immunespecific antibody by immunoaffinity chromatography. Purification ofimmunoglobulins is discussed, for example, by D. Wilkinson (TheScientist, published by The Scientist, Inc., Philadelphia Pa., Vol. 14,No. 8 (Apr. 17, 2000), pp. 25-28).

[0224] II. Monoclonal Antibodies

[0225] The term “monoclonal antibody” (MAb) or “monoclonal antibodycomposition”, as used herein, refers to a population of antibodymolecules that contain only one molecular species of antibody moleculeconsisting of a unique light chain gene product and a unique heavy chaingene product. In particular, the complementarity determining regions(CDRs) of the monoclonal antibody are identical in all the molecules ofthe population. MAbs thus contain an antigen binding site capable ofimmunoreacting with a particular epitope of the antigen characterized bya unique binding affinity for it.

[0226] Monoclonal antibodies can be prepared using hybridoma methods,such as those described by Kohler and Milstein, (1975. Nature 256: 495).In a hybridoma method, a mouse, hamster, or other appropriate hostanimal, is typically immunized with an immunizing agent to elicitlymphocytes that produce or are capable of producing antibodies thatwill specifically bind to the immunizing agent. Alternatively, thelymphocytes can be immunized in vitro.

[0227] The immunizing agent will typically include the protein antigen,a fragment thereof or a fusion protein thereof. Generally, eitherperipheral blood lymphocytes are used if cells of human origin aredesired, or spleen cells or lymph node cells are used if non-humanmammalian sources are desired. The lymphocytes are then fused with animmortalized cell line using a suitable fusing agent, such aspolyethylene glycol, to form a hybridoma cell (Goding, MONOCLONALANTIBODIES: PRINCIPLES AND PRACTICE, Academic Press, (1986) pp. 59-103).Immortalized cell lines are usually transformed mammalian cells,particularly myeloma cells of rodent, bovine and human origin. Usually,rat or mouse myeloma cell lines are employed. The hybridoma cells can becultured in a suitable culture medium that preferably contains one ormore substances that inhibit the growth or survival of the unfused,immortalized cells. For example, if the parental cells lack the enzymehypoxanthine guanine phosphoribosyl transferase (HGPRT or HPRT), theculture medium for the hybridomas typically will include hypoxanthine,aminopterin, and thymidine (“HAT medium”), which substances prevent thegrowth of HGPRT-deficient cells.

[0228] Preferred immortalized cell lines are those that fuseefficiently, support stable high level expression of antibody by theselected antibody-producing cells, and are sensitive to a medium such asHAT medium. More preferred immortalized cell lines are murine myelomalines, which can be obtained, for instance, from the Salk Institute CellDistribution Center, San Diego, Calif. and the American Type CultureCollection, Manassas, Va. Human myeloma and mouse-human heteromyelomacell lines also have been described for the production of humanmonoclonal antibodies (Kozbor, 1984. J. Immunol. 133: 3001; Brodeur, etal., MONOCLONAL ANTIBODY PRODUCTION TECHNIQUES AND APPLICATIONS, MarcelDekker, Inc., New York, (1987) pp. 51-63).

[0229] The culture medium in which the hybridoma cells are cultured canthen be assayed for the presence of monoclonal antibodies directedagainst the antigen. Preferably, the binding specificity of monoclonalantibodies produced by the hybridoma cells is determined byimmunoprecipitation or by an in vitro binding assay, such asradioimmunoassay (RIA) or enzyme-linked immunoabsorbent assay (ELISA).Such techniques and assays are known in the art. The binding affinity ofthe monoclonal antibody can, for example, be determined by the Scatchardanalysis of Munson and Pollard, (1980. Anal. Biochem. 107: 220).Preferably, antibodies having a high degree of specificity and a highbinding affinity for the target antigen are isolated.

[0230] After the desired hybridoma cells are identified, the clones canbe subcloned by limiting dilution procedures and grown by standardmethods. Suitable culture media for this purpose include, for example,Dulbecco's Modified Eagle's Medium and RPMI-1640 medium. Alternatively,the hybridoma cells can be grown in vivo as ascites in a mammal.

[0231] The monoclonal antibodies secreted by the subclones can beisolated or purified from the culture medium or ascites fluid byconventional immunoglobulin purification procedures such as, forexample, protein A-Sepharose, hydroxylapatite chromatography, gelelectrophoresis, dialysis, or affinity chromatography.

[0232] The monoclonal antibodies can also be made by recombinant DNAmethods, such as those described in U.S. Pat. No. 4,816,567. DNAencoding the monoclonal antibodies of the invention can be readilyisolated and sequenced using conventional procedures (e.g., by usingoligonucleotide probes that are capable of binding specifically to genesencoding the heavy and light chains of murine antibodies). The hybridomacells of the invention serve as a preferred source of such DNA. Onceisolated, the DNA can be placed into expression vectors, which are thentransfected into host cells such as simian COS cells, Chinese hamsterovary (CHO) cells, or myeloma cells that do not otherwise produceimmunoglobulin protein, to obtain the synthesis of monoclonal antibodiesin the recombinant host cells. The DNA also can be modified, forexample, by substituting the coding sequence for human heavy and lightchain constant domains in place of the homologous murine sequences (U.S.Pat. No. 4,816,567; Morrison, 1994. Nature 368: 812-813) or bycovalently joining to the immunoglobulin coding sequence all or part ofthe coding sequence for a non-immunoglobulin polypeptide. Such anon-immunoglobulin polypeptide can be substituted for the constantdomains of an antibody of the invention, or can be substituted for thevariable domains of one antigen-combining site of an antibody of theinvention to create a chimeric bivalent antibody.

[0233] III. Humanized Antibodies

[0234] The antibodies directed against the protein antigens of theinvention can further comprise humanized antibodies or human antibodies.These antibodies are suitable for administration to humans withoutengendering an immune response by the human against the administeredimmunoglobulin. Humanized forms of antibodies are chimericimmunoglobulins, immunoglobulin chains or fragments thereof (such as Fv,Fab, Fab′, F(ab′)₂ or other antigen-binding subsequences of antibodies)that are principally comprised of the sequence of a humanimmunoglobulin, and contain minimal sequence derived from a non-humanimmunoglobulin. Humanization can be performed following the method ofWinter and co-workers (Jones, et al., 1986. Nature, 321: 522-525;Riechmann, et al., 1988. Nature 332: 323-327; Verhoeyen, et al., 1988.Science, 239: 1534-1536); by substituting rodent CDRs or CDR sequencesfor the corresponding sequences of a human antibody. (see, e.g., U.S.Pat. No. 5,225,539.). In some instances, Fv framework residues of thehuman immunoglobulin are replaced by corresponding non-human residues.Humanized antibodies can also comprise residues which are found neitherin the recipient antibody nor in the imported CDR or frameworksequences. In general, the humanized antibody will comprisesubstantially all of at least one, and typically two, variable domains,in which all or substantially all of the CDR regions correspond to thoseof a non-human immunoglobulin and all or substantially all of theframework regions are those of a human immunoglobulin consensussequence. The humanized antibody optimally also will comprise at least aportion of an immunoglobulin constant region (Fc), typically that of ahuman immunoglobulin (Jones, et al., 1986, supra; Riechmann, et al.,1988, supra; Presta, 1992. Curr. Op. Struct. Biol., 2: 593-596).

[0235] IV. Human Antibodies

[0236] Fully human antibodies relate to antibody molecules in whichessentially the entire sequences of both the light chain and the heavychain, including the CDRs, arise from human genes. Such antibodies aretermed “human antibodies”, or “fully human antibodies” herein. Humanmonoclonal antibodies can be prepared by the trioma technique; the humanB-cell hybridoma technique (see, e.g., Kozbor, et al., 1983. ImmunolToday 4: 72) and the EBV hybridoma technique to produce human monoclonalantibodies (see, e.g., Cole, et al., 1985. In: MONOCLONAL ANTIBODIES ANDCANCER THERAPY, Alan R. Liss, Inc., pp. 77-96). Human monoclonalantibodies may be utilized in the practice of the present invention andmay be produced by using human hybridomas (see, e.g., Cote, et al.,1983. Proc. Natl. Acad. Sci. USA 80: 2026-2030) or by transforming humanB-cells with Epstein Barr Virus in vitro (see, e.g., Cole, et al.,1985., supra).

[0237] In addition, human antibodies can also be produced usingadditional techniques, including phage display libraries (Hoogenboom andWinter, 1991. J. Mol. Biol. 227: 381; Marks, et al., J. Mol. Biol.222:). Similarly, human antibodies can be made by introducing humanimmunoglobulin loci into transgenic animals, e.g., mice in which theendogenous immunoglobulin genes have been partially or completelyinactivated. Upon challenge, human antibody production is observed,which closely resembles that seen in humans in all respects, includinggene rearrangement, assembly, and antibody repertoire. This approach isdescribed, for example, in U.S. Pat. Nos. 5,545,807; 5,545,806;5,569,825; 5,625,126; 5,633,425; 5,661,016, and Marks, et al. (1992.Bio/Technology 10: 779-783); Lonberg, et al. (1994. Nature 368:856-859); Morrison (1994. Nature 368: 812-813); Fishwild, et al, (1996.Nature Biotech. 14: 845-851); Neuberger (1996. Nature Biotech. 14: 826);and Lonberg and Huszar (1995. International Rev. Immunol. 13: 65-93).

[0238] Human antibodies may additionally be produced using transgenicnonhuman animals which are modified so as to produce fully humanantibodies rather than the animal's endogenous antibodies in response tochallenge by an antigen. See, PCT Publication WO94/02602. The endogenousgenes encoding the heavy and light immunoglobulin chains in the nonhumanhost have been incapacitated, and active loci encoding human heavy andlight chain immunoglobulins are inserted into the host's genome. Thehuman genes are incorporated, for example, using yeast artificialchromosomes containing the requisite human DNA segments. An animal whichprovides all the desired modifications is then obtained as progeny bycrossbreeding intermediate transgenic animals containing fewer than thefull complement of the modifications. The preferred embodiment of such anonhuman animal is a mouse, and is termed the Xenomouse™ as disclosed inPCT Publications WO 96/33735 and WO 96/34096. This animal produces Bcells which secrete fully human immunoglobulins. The antibodies can beobtained directly from the animal after immunization with an immunogenof interest, as, for example, a preparation of a polyclonal antibody, oralternatively from immortalized B cells derived from the animal, such ashybridomas producing monoclonal antibodies. Additionally, the genesencoding the immunoglobulins with human variable regions can berecovered and expressed to obtain the antibodies directly, or can befurther modified to obtain analogs of antibodies such as, for example,single chain Fv molecules.

[0239] An example of a method of producing a nonhuman host, exemplifiedas a mouse, lacking expression of an endogenous immunoglobulin heavychain is disclosed in U.S. Pat. No. 5,939,598. It can be obtained by amethod including deleting the J segment genes from at least oneendogenous heavy chain locus in an embryonic stem cell to preventrearrangement of the locus and to prevent formation of a transcript of arearranged immunoglobulin heavy chain locus, the deletion being effectedby a targeting vector containing a gene encoding a selectable marker;and producing from the embryonic stem cell a transgenic mouse whosesomatic and germ cells contain the gene encoding the selectable marker.

[0240] A method for producing an antibody of interest, such as a humanantibody, is disclosed in U.S. Pat. No. 5,916,771. It includesintroducing an expression vector that contains a nucleotide sequenceencoding a heavy chain into one mammalian host cell in culture,introducing an expression vector containing a nucleotide sequenceencoding a light chain into another mammalian host cell, and fusing thetwo cells to form a hybrid cell. The hybrid cell expresses an antibodycontaining the heavy chain and the light chain.

[0241] In a further improvement on this procedure, a method foridentifying a clinically relevant epitope on an immunogen, and acorrelative method for selecting an antibody that bindsimmunospecifically to the relevant epitope with high affinity, aredisclosed in PCT publication WO 99/53049.

[0242] V. Fa_(b) Fragments and Single Chain Antibodies

[0243] According to the invention, techniques can be adapted for theproduction of single-chain antibodies specific to an antigenic proteinof the invention (see, e.g., U.S. Pat. No. 4,946,778). In addition,methods can be adapted for the construction of F_(ab) expressionlibraries (see, e.g., Huse, et al., 1989 Science 246: 1275-1281) toallow rapid and effective identification of monoclonal F_(ab) fragmentswith the desired specificity for a protein or derivatives, fragments,analogs or homologs thereof. Antibody fragments that contain theidiotypes to a protein antigen may be produced by techniques known inthe art including, but not limited to: (i) an F_((ab′)2) fragmentproduced by pepsin digestion of an antibody molecule; (ii) an F_(ab)fragment generated by reducing the disulfide bridges of an F_((ab′)2)fragment; (iii) an F_(ab) fragment generated by the treatment of theantibody molecule with papain and a reducing agent; and (iv) F_(v)fragments.

[0244] VI. Bispecific Antibodies

[0245] Bispecific antibodies are monoclonal, preferably human orhumanized, antibodies that have binding specificities for at least twodifferent antigens. In the present case, one of the bindingspecificities is for an antigenic protein of the invention. The secondbinding target is any other antigen, and advantageously is acell-surface protein or receptor or receptor subunit.

[0246] Methods for making bispecific antibodies are known in the art.Traditionally, the recombinant production of bispecific antibodies isbased on the co-expression of two immunoglobulin heavy-chain/light-chainpairs, where the two heavy chains have different specificities (see,e.g., Milstein and Cuello, 1983. Nature 305: 537-539). Because of therandom assortment of immunoglobulin heavy and light chains, thesehybridomas (i.e., quadromas) produce a potential mixture of tendifferent antibody molecules, of which only one has the correctbispecific structure. The purification of the correct molecule isusually accomplished by affinity chromatography steps. Similarprocedures are disclosed in PCT Publication WO 93/08829 (published May13, 1993); Traunecker, et al., (1991. EMBO J., 10: 3655-3659).

[0247] Antibody variable domains with the desired binding specificities(antibody-antigen combining sites) can be fused to immunoglobulinconstant domain sequences. The fusion preferably is with animmunoglobulin heavy-chain constant domain, comprising at least part ofthe hinge, CH2, and CH3 regions. It is preferred to have the firstheavy-chain constant region (CH1) containing the site necessary forlight-chain binding present in at least one of the fusions. DNAsencoding the immunoglobulin heavy-chain fusions and, if desired, theimmunoglobulin light chain, are inserted into separate expressionvectors, and are co-transfected into a suitable host organism. Forfurther details of generating bispecific antibodies (see, e.g., Suresh,et al., 1986. Meth. Enzymology 121: 210).

[0248] According to another approach described in PCT Publication WO96/27011, the interface between a pair of antibody molecules can beengineered to maximize the percentage of heterodimers which arerecovered from recombinant cell culture. The preferred interfacecomprises at least a part of the CH3 region of an antibody constantdomain. In this method, one or more small amino acid side chains fromthe interface of the first antibody molecule are replaced with largerside chains (e.g., tyrosine or tryptophan). Compensatory “cavities” ofidentical or similar size to the large side chain(s) are created on theinterface of the second antibody molecule by replacing large amino acidside chains with smaller ones (e.g. alanine or threonine). This providesa mechanism for increasing the yield of the heterodimer over otherunwanted end-products such as homodimers.

[0249] Bispecific antibodies can be prepared as full length antibodiesor antibody fragments (e.g. F_((ab′)2) bispecific antibodies).Techniques for generating bispecific antibodies from antibody fragmentshave been described in the literature. For example, bispecificantibodies can be prepared using chemical linkage. Brennan, et al.(1985. Science 229: 81) describe a procedure wherein intact antibodiesare proteolytically cleaved to generate F_((ab′)2) fragments. Thesefragments are reduced in the presence of the dithiol complexing agentsodium arsenite to stabilize vicinal dithiols and prevent intermoleculardisulfide formation. The Fab′ fragments generated are then converted tothionitrobenzoate (TNB) derivatives. One of the Fab′-TNB derivatives isthen reconverted to the Fab′-thiol by reduction with mercaptoethylamineand is mixed with an equimolar amount of the other Fab′-TNB derivativeto form the bispecific antibody. The bispecific antibodies produced canbe used as agents for the selective immobilization of enzymes.

[0250] Additionally, Fab′ fragments can be directly recovered from E.coli and chemically coupled to form bispecific antibodies. Shalaby, etal. (1992. J. Exp. Med. 175: 217-225) describe the production of a fullyhumanized bispecific antibody F_((ab′)2) molecule. Each Fab′ fragmentwas separately secreted from E. coli and subjected to directed chemicalcoupling in vitro to form the bispecific antibody. The bispecificantibody thus formed was able to bind to cells overexpressing the ErbB2receptor and normal human T cells, as well as trigger the lytic activityof human cytotoxic lymphocytes against human breast tumor targets.

[0251] Various techniques for making and isolating bispecific antibodyfragments directly from recombinant cell culture have also beendescribed. For example, bispecific antibodies have been produced usingleucine zippers (Kostelny, et al., 1992. J. Immunol. 148(5): 1547-1553).The leucine zipper peptides from the Fos and Jun proteins were linked tothe Fab′ portions of two different antibodies by gene fusion. Theantibody homodimers were reduced at the hinge region to form monomersand then re-oxidized to form the antibody beterodimers. This method canalso be utilized for the production of antibody homodimers. The“diabody” technology described by Hollinger, et al. (1993. Proc. Natl.Acad. Sci. USA 90: 6444-6448) has provided an alternative mechanism formaking bispecific antibody fragments. The fragments comprise aheavy-chain variable domain (V_(H)) connected to a light-chain variabledomain (V_(L)) by a linker which is too short to allow pairing betweenthe two domains on the same chain. Accordingly, the V_(H) and V_(L)domains of one fragment are forced to pair with the complementary V_(L)and V_(H) domains of another fragment, thereby forming twoantigen-binding sites. Another strategy for making bispecific antibodyfragments by the use of single-chain Fv (sFv) dimers has also beenreported (Gruber, et al., 1994. J. Immunol. 152: 5368).

[0252] Antibodies with more than two valencies are contemplated. Forexample, trispecific antibodies can be prepared (Tutt, et al., 1991. J.Immunol. 147: 60).

[0253] Exemplary bispecific antibodies can bind to two differentepitopes, at least one of which originates in the protein antigen of theinvention. Alternatively, an anti-antigenic arm of an immunoglobulinmolecule can be combined with an arm which binds to a triggeringmolecule on a leukocyte such as a T-cell receptor molecule (e.g., CD2,CD3, CD28, or B7), or Fc receptors for IgG (Fc R), such as Fc RI (CD64),Fc RII (CD32) and Fc RIII (CD16) so as to focus cellular defensemechanisms to the cell expressing the particular antigen. Bispecificantibodies can also be used to direct cytotoxic agents to cells whichexpress a particular antigen. These antibodies possess anantigen-binding arm and an arm which binds a cytotoxic agent or aradionuclide chelator, such as EOTUBE, DPTA, DOTA, or TETA. Anotherbispecific antibody of interest binds the protein antigen describedherein and further binds tissue factor (TF).

[0254] VII. Heteroconjugate Antibodies

[0255] Heteroconjugate antibodies are also within the scope of thepresent invention. Heteroconjugate antibodies are composed of twocovalently joined antibodies. Such antibodies have, for example, beenproposed to target immune system cells to unwanted cells (U.S. Pat. No.4,676,980), and for treatment of HIV infection (WO 91/00360; WO92/200373; EP 03089). It is contemplated that the antibodies can beprepared in vitro using known methods in synthetic protein chemistry,including those involving cross-linking agents. For example,immunotoxins can be constructed using a disulfide exchange reaction orby forming a thioether bond. Examples of suitable reagents for thispurpose include iminothiolate and methyl-4-mercaptobutyrimidate andthose disclosed, e.g., in U.S. Pat. No. 4,676,980.

[0256] VIII. Effector Function Engineering

[0257] It can be desirable to modify the antibody of the invention withrespect to effector function, so as to enhance, e.g., the effectivenessof the antibody in treating cancer. For example, cysteine residue(s) canbe introduced into the Fc region, thereby allowing interchain disulfidebond formation in this region. The homodimeric antibody thus generatedcan have improved internalization capability and/or increasedcomplement-mediated cell killing and antibody-dependent cellularcytotoxicity (ADCC). See, e.g., Caron, et al., 1992. J. Exp Med., 176:1191-1195; Shopes, 1992. J. Immunol. 148: 2918-2922. Homodimericantibodies with enhanced anti-tumor activity can also be prepared usingheterobifunctional cross-linkers as described by Wolff, et al. (1993.Cancer Res. 53: 2560-2565). Alternatively, an antibody can be engineeredthat has dual Fc regions and can thereby have enhanced complement lysisand ADCC capabilities (see, e.g., Stevenson, et al., 1989. Anti-CancerDrug Design 3: 219-230).

[0258] IX. Immunoconjugates

[0259] The invention also pertains to immunoconjugates comprising anantibody conjugated to a cytotoxic agent such as a chemotherapeuticagent, toxin (e.g., an enzymatically active toxin of bacterial, fungal,plant, or animal origin, or fragments thereof), or a radioactive isotope(i.e., a radioconjugate).

[0260] Chemotherapeutic agents useful in the generation of suchimmunoconjugates have been described above. Enzymatically active toxinsand fragments thereof that can be used include diphtheria A chain,nonbinding active fragments of diphtheria toxin, exotoxin A chain (fromPseudomonas aeruginosa), ricin A chain, abrin A chain, modeccin A chain,alpha-sarcin, Aleurites fordii proteins, dianthin proteins, Phytolacaamericana proteins (PAPI, PAPII, and PAP-S), momordica charantiainhibitor, curcin, crotin, sapaonaria officinalis inhibitor, gelonin,mitogellin, restrictocin, phenomycin, enomycin, and the tricothecenes. Avariety of radionuclides are available for the production ofradioconjugated antibodies. Examples include ²¹²Bi, ¹³¹I, ¹³¹In, ⁹⁰Y,and ¹⁸⁶Re.

[0261] Conjugates of the antibody and cytotoxic agent are made using avariety of bifunctional protein-coupling agents such asN-succinimidyl-3-(2-pyridyldithiol) propionate (SPDP), iminothiolane(IT), bifunctional derivatives of imidoesters (such as dimethyladipimidate HCL), active esters (such as disuccinimidyl suberate),aldehydes (such as glutareldehyde), bis-azido compounds (such as bis(p-azidobenzoyl) hexanediamine), bis-diazonium derivatives (such asbis-(p-diazoniumbenzoyl)-ethylenediamine), diisocyanates (such astolyene 2,6-diisocyanate), and bis-active fluorine compounds (such as1,5-difluoro-2,4-dinitrobenzene). For example, a ricin immunotoxin canbe prepared as described by Vitetta, et al. (1987. Science 238:1098).Carbon-14-labeled, 1-isothiocyanatobenzyl-3-methyldiethylenetriaminepentaacetic acid (MX-DTPA) is an exemplary chelating agent forconjugation of radionucleotide to the antibody. See, PCT PublicationWO94/11026.

[0262] In another embodiment, the antibody can be conjugated to a“receptor” (such streptavidin) for utilization in tumor pretargetingwherein the antibody-receptor conjugate is administered to the patient,followed by removal of unbound conjugate from the circulation using aclearing agent and then administration of a “ligand” (e.g., avidin) thatis in turn conjugated to a cytotoxic agent.

[0263] MEMX Recombinant Expression Vectors and Host Cells

[0264] Another aspect of the invention pertains to vectors, preferablyexpression vectors, containing a nucleic acid encoding an MEMX protein,or derivatives, fragments, analogs or homologs thereof. As used herein,the term “vector” refers to a nucleic acid molecule capable oftransporting another nucleic acid to which it has been linked. One typeof vector is a “plasmid”, which refers to a circular double stranded DNAloop into which additional DNA segments can be ligated. Another type ofvector is a viral vector, wherein additional DNA segments can be ligatedinto the viral genome. Certain vectors are capable of autonomousreplication in a host cell into which they are introduced (e.g.,bacterial vectors having a bacterial origin of replication and episomalmammalian vectors). Other vectors (e.g., non-episomal mammalian vectors)are integrated into the genome of a host cell upon introduction into thehost cell, and thereby are replicated along with the host genome.Moreover, certain vectors are capable of directing the expression ofgenes to which they are operatively-linked. Such vectors are referred toherein as “expression vectors”. In general, expression vectors ofutility in recombinant DNA techniques are often in the form of plasmids.In the present specification, “plasmid” and “vector” can be usedinterchangeably as the plasmid is the most commonly used form of vector.However, the invention is intended to include such other forms ofexpression vectors, such as viral vectors (e.g., replication defectiveretroviruses, adenoviruses and adeno-associated viruses), which serveequivalent functions.

[0265] The recombinant expression vectors of the invention comprise anucleic acid of the invention in a form suitable for expression of thenucleic acid in a host cell, which means that the recombinant expressionvectors include one or more regulatory sequences, selected on the basisof the host cells to be used for expression, that is operatively-linkedto the nucleic acid sequence to be expressed. Within a recombinantexpression vector, “operably-linked” is intended to mean that thenucleotide sequence of interest is linked to the regulatory sequence(s)in a manner that allows for expression of the nucleotide sequence (e.g.,in an in vitro transcription/translation system or in a host cell whenthe vector is introduced into the host cell).

[0266] The term “regulatory sequence” is intended to includes promoters,enhancers and other expression control elements (e.g., polyadenylationsignals). Such regulatory sequences are described, for example, inGoeddel, GENE EXPRESSION TECHNOLOGY: METHODS IN ENZYMOLOGY 185, AcademicPress, San Diego, Calif. (1990). Regulatory sequences include those thatdirect constitutive expression of a nucleotide sequence in many types ofhost cell and those that direct expression of the nucleotide sequenceonly in certain host cells (e.g., tissue-specific regulatory sequences).It will be appreciated by those skilled in the art that the design ofthe expression vector can depend on such factors as the choice of thehost cell to be transformed, the level of expression of protein desired,etc. The expression vectors of the invention can be introduced into hostcells to thereby produce proteins or peptides, including fusion proteinsor peptides, encoded by nucleic acids as described herein (e.g., MEMXproteins, mutant forms of MEMX proteins, fusion proteins, etc.).

[0267] The recombinant expression vectors of the invention can bedesigned for expression of MEMX proteins in prokaryotic or eukaryoticcells. For example, MEMX proteins can be expressed in bacterial cellssuch as Escherichia coli, insect cells (using baculovirus expressionvectors) yeast cells or mammalian cells. Suitable host cells arediscussed further in Goeddel, GENE EXPRESSION TECHNOLOGY: METHODS INENZYMOLOGY 185, Academic Press, San Diego, Calif. (1990). Alternatively,the recombinant expression vector can be transcribed and translated invitro, for example using T7 promoter regulatory sequences and T7polymerase.

[0268] Expression of proteins in prokaryotes is most often carried outin Escherichia coli with vectors containing constitutive or induciblepromoters directing the expression of either fusion or non-fusionproteins. Fusion vectors add a number of amino acids to a proteinencoded therein, usually to the amino terminus of the recombinantprotein. Such fusion vectors typically serve three purposes: (i) toincrease expression of recombinant protein; (ii) to increase thesolubility of the recombinant protein; and (iii) to aid in thepurification of the recombinant protein by acting as a ligand inaffinity purification. Often, in fusion expression vectors, aproteolytic cleavage site is introduced at the junction of the fusionmoiety and the recombinant protein to enable separation of therecombinant protein from the fusion moiety subsequent to purification ofthe fusion protein. Such enzymes, and their cognate recognitionsequences, include Factor Xa, thrombin and enterokinase. Typical fusionexpression vectors include pGEX (Pharmacia Biotech Inc; Smith andJohnson, 1988. Gene 67: 31-40), pMAL (New England Biolabs, Beverly,Mass.) and pRIT5 (Pharmacia, Piscataway, N.J.) that fuse glutathioneS-transferase (GST), maltose E binding protein, or protein A,respectively, to the target recombinant protein.

[0269] Examples of suitable inducible non-fusion E. coli expressionvectors include pTrc (Amrann et al., (1988) Gene 69:301-315) and pET 11d(Studier et al., GENE EXPRESSION TECHNOLOGY: METHODS IN ENZYMOLOGY 185,Academic Press, San Diego, Calif. (1990) 60-89).

[0270] One strategy to maximize recombinant protein expression in E.coli is to express the protein in a host bacteria with an impairedcapacity to proteolytically cleave the recombinant protein. See, e.g.,Gottesman, GENE EXPRESSION TECHNOLOGY: METHODS IN ENZYMOLOGY 185,Academic Press, San Diego, Calif. (1990) 119-128. Another strategy is toalter the nucleic acid sequence of the nucleic acid to be inserted intoan expression vector so that the individual codons for each amino acidare those preferentially utilized in E. coli (see, e.g., Wada, et al.,1992. Nucl. Acids Res. 20: 2111-2118). Such alteration of nucleic acidsequences of the invention can be carried out by standard DNA synthesistechniques.

[0271] In another embodiment, the MEMX expression vector is a yeastexpression vector. Examples of vectors for expression in yeastSaccharomyces cerivisae include pYepSec1 (Baldari, et al., 1987. EMBO J.6: 229-234), pMFa (Kurjan and Herskowitz, 1982. Cell 30: 933-943),pJRY88 (Schultz et al., 1987. Gene 54: 113-123), pYES2 (InvitrogenCorporation, San Diego, Calif.), and picZ (InVitrogen Corp, San Diego,Calif.).

[0272] Alternatively, MEMX can be expressed in insect cells usingbaculovirus expression vectors. Baculovirus vectors available forexpression of proteins in cultured insect cells (e.g., SF9 cells)include the pAc series (Smith, et al., 1983. Mol. Cell. Biol. 3:2156-2165) and the pVL series (Lucklow and Summers, 1989. Virology 170:31-39).

[0273] In yet another embodiment, a nucleic acid of the invention isexpressed in mammalian cells using a mammalian expression vector.Examples of mammalian expression vectors include pCDM8 (Seed, 1987.Nature 329: 840) and pMT2PC (Kaufman, et al., 1987. EMBO J. 6: 187-195).When used in mammalian cells, the expression vector's control functionsare often provided by viral regulatory elements. For example, commonlyused promoters are derived from polyoma, adenovirus 2, cytomegalovirus,and simian virus 40. For other suitable expression systems for bothprokaryotic and eukaryotic cells see, e.g., Chapters 16 and 17 ofSambrook, et al., MOLECULAR CLONING: A LABORATORY MANUAL. 2nd ed., ColdSpring Harbor Laboratory, Cold Spring Harbor Laboratory Press, ColdSpring Harbor, N.Y., 1989.

[0274] In another embodiment, the recombinant mammalian expressionvector is capable of directing expression of the nucleic acidpreferentially in a particular cell type (e.g., tissue-specificregulatory elements are used to express the nucleic acid).Tissue-specific regulatory elements are known in the art. Non-limitingexamples of suitable tissue-specific promoters include the albuminpromoter (liver-specific; Pinkert, et al., 1987. Genes Dev. 1: 268-277),lymphoid-specific promoters (Calame and Eaton, 1988. Adv. Immunol. 43:235-275), in particular promoters of T cell receptors (Winoto andBaltimore, 1989. EMBO J. 8: 729-733) and immunoglobulins (Banerji, etal., 1983. Cell 33: 729-740; Queen and Baltimore, 1983. Cell 33:741-748), neuron-specific promoters (e.g., the neurofilament promoter;Byrne and Ruddle, 1989. Proc. Natl. Acad. Sci. USA 86: 5473-5477),pancreas-specific promoters (Edlund, et al., 1985. Science 230:912-916), and mammary gland-specific promoters (e.g., milk wheypromoter; U.S. Pat. No. 4,873,316 and European Application PublicationNo. 264,166). Developmentally-regulated promoters are also encompassed,e.g., the murine hox promoters (Kessel and Gruss, 1990. Science 249:374-379) and the α-fetoprotein promoter (Campes and Tilghman, 1989.Genes Dev. 3: 537-546).

[0275] The invention further provides a recombinant expression vectorcomprising a DNA molecule of the invention cloned into the expressionvector in an antisense orientation. That is, the DNA molecule isoperatively-linked to a regulatory sequence in a manner that allows forexpression (by transcription of the DNA molecule) of an RNA moleculethat is antisense to MEMX mRNA. Regulatory sequences operatively linkedto a nucleic acid cloned in the antisense orientation can be chosen thatdirect the continuous expression of the antisense RNA molecule in avariety of cell types, for instance viral promoters and/or enhancers, orregulatory sequences can be chosen that direct constitutive, tissuespecific or cell type specific expression of antisense RNA. Theantisense expression vector can be in the form of a recombinant plasmid,phagemi d or attenuated virus in which antisense nucleic acids areproduced under the control of a high efficiency regulatory region, theactivity of which can be determined by the cell type into which thevector is introduced. For a discussion of the regulation of geneexpression using antisense genes see, e.g., Weintraub, et al.,“Antisense RNA as a molecular tool for genetic analysis,” Reviews-Trendsin Genetics, Vol. 1(1) 1986.

[0276] Another aspect of the invention pertains to host cells into whicha recombinant expression vector of the invention has been introduced.The terms “host cell” and “recombinant host cell” are usedinterchangeably herein. It is understood that such terms refer not onlyto the particular subject cell but also to the progeny or potentialprogeny of such a cell. Because certain modifications may occur insucceeding generations due to either mutation or environmentalinfluences, such progeny may not, in fact, be identical to the parentcell, but are still included within the scope of the term as usedherein.

[0277] A host cell can be any prokaryotic or eukaryotic cell. Forexample, MEMX protein can be expressed in bacterial cells such as E.coli, insect cells, yeast or mammalian cells (e.g., Chinese hamsterovary cells (CHO) or COS cells). Other suitable host cells are known tothose skilled in the art.

[0278] Vector DNA can be introduced into prokaryotic or eukaryotic cellsvia conventional transformation or transfection techniques. As usedherein, the terms “transformation” and “transfection” are intended torefer to a variety of art-recognized techniques for introducing foreignnucleic acid (e.g., DNA) into a host cell, including calcium phosphateor calcium chloride co-precipitation, DEAE-dextran-mediatedtransfection, lipofection, or electroporation. Suitable methods fortransforming or transfecting host cells can be found in Sambrook, et al.(MOLECULAR CLONING: A LABORATORY MANUAL. 2nd ed., Cold Spring HarborLaboratory, Cold Spring Harbor Laboratory Press, Cold Spring Harbor,N.Y., 1989), and other laboratory manuals.

[0279] For stable transfection of mammalian cells, it is known that,depending upon the expression vector and transfection technique used,only a small fraction of cells may integrate the foreign DNA into theirgenome. In order to identify and select these integrants, a gene thatencodes a selectable marker (e.g., resistance to antibiotics) isgenerally introduced into the host cells along with the gene ofinterest. Various selectable markers include those that conferresistance to drugs, such as G418, hygromycin and methotrexate. Nucleicacid encoding a selectable marker can be introduced into a host cell onthe same vector as that encoding MEMX or can be introduced on a separatevector. Cells stably transfected with the introduced nucleic acid can beidentified by drug selection (e.g., cells that have incorporated theselectable marker gene will survive, while the other cells die).

[0280] A host cell of the invention, such as a prokaryotic or eukaryotichost cell in culture, can be used to produce (i.e., express) MEMXprotein. Accordingly, the invention further provides methods forproducing MEMX protein using the host cells of the invention. In oneembodiment, the method comprises culturing the host cell of invention(into which a recombinant expression vector encoding MEMX protein hasbeen introduced) in a suitable medium such that MEMX protein isproduced. In another embodiment, the method further comprises isolatingMEMX protein from the medium or the host cell.

[0281] Transgenic MEMX Animals

[0282] The host cells of the invention can also be used to producenon-human transgenic animals. For example, in one embodiment, a hostcell of the invention is a fertilized oocyte or an embryonic stem cellinto which MEMX protein-coding sequences have been introduced. Such hostcells can then be used to create non-human transgenic animals in whichexogenous MEMX sequences have been introduced into their genome orhomologous recombinant animals in which endogenous MEMX sequences havebeen altered. Such animals are useful for studying the function and/oractivity of MEMX protein and for identifying and/or evaluatingmodulators of MEMX protein activity. As used herein, a “transgenicanimal” is a non-human animal, preferably a mammal, more preferably arodent such as a rat or mouse, in which one or more of the cells of theanimal includes a transgene. Other examples of transgenic animalsinclude non-human primates, sheep, dogs, cows, goats, chickens,amphibians, etc. A transgene is exogenous DNA that is integrated intothe genome of a cell from which a transgenic animal develops and thatremains in the genome of the mature animal, thereby directing theexpression of an encoded gene product in one or more cell types ortissues of the transgenic animal. As used herein, a “homologousrecombinant animal” is a non-human animal, preferably a mammal, morepreferably a mouse, in which an endogenous MEMX gene has been altered byhomologous recombination between the endogenous gene and an exogenousDNA molecule introduced into a cell of the animal, e.g., an embryoniccell of the animal, prior to development of the animal.

[0283] A transgenic animal of the invention can be created byintroducing MEMX-encoding nucleic acid into the male pronuclei of afertilized oocyte (e.g., by microinjection, retroviral infection) andallowing the oocyte to develop in a pseudopregnant female foster animal.The human MEMX cDNA sequences of SEQ ID NO:1, 3, 5, 7, 9, 11, 13, or 15,can be introduced as a transgene into the genome of a non-human animal.Alternatively, a non-human homologue of the human MEMX gene, such as amouse MEMX gene, can be isolated based on hybridization to the humanMEMX cDNA (described further supra) and used as a transgene. Intronicsequences and polyadenylation signals can also be included in thetransgene to increase the efficiency of expression of the transgene. Atissue-specific regulatory sequence(s) can be operably-linked to theMEMX transgene to direct expression of MEMX protein to particular cells.Methods for generating transgenic animals via embryo manipulation andmicroinjection, particularly animals such as mice, have becomeconventional in the art and are described, for example, in U.S. Pat.Nos. 4,736,866; 4,870,009; and 4,873,191; and Hogan, 1986. In:MANIPULATING THE MOUSE EMBRYO, Cold Spring Harbor Laboratory Press, ColdSpring Harbor, N.Y. Similar methods are used for production of othertransgenic animals. A transgenic founder animal can be identified basedupon the presence of the MEMX transgene in its genome and/or expressionof MEMX mRNA in tissues or cells of the animals. A transgenic founderanimal can then be used to breed additional animals carrying thetransgene. Moreover, transgenic animals carrying a transgene-encodingMEMX protein can further be bred to other transgenic animals carryingother transgenes.

[0284] To create a homologous recombinant animal, a vector is preparedwhich contains at least a portion of an MEMX gene into which a deletion,addition or substitution has been introduced to thereby alter, e.g.,functionally disrupt, the MEMX gene. The MEMX gene can be a human gene(e.g., the cDNA of SEQ ID NO:1, 3, 5, 7, 9, 11, 13, or 15), but morepreferably, is a non-human homologue of a human MEMX gene. For example,a mouse homologue of human MEMX gene of SEQ ID NO:1, 3, 5, 7, 9, 11, 13,or 15, can be used to construct a homologous recombination vectorsuitable for altering an endogenous MEMX gene in the mouse genome. Inone embodiment, the vector is designed such that, upon homologousrecombination, the endogenous MEMX gene is functionally disrupted (i.e.,no longer encodes a functional protein; also referred to as a “knockout” vector).

[0285] Alternatively, the vector can be designed such that, uponhomologous recombination, the endogenous MEMX gene is mutated orotherwise altered but still encodes functional protein (e.g., theupstream regulatory region can be altered to thereby alter theexpression of the endogenous MEMX protein). In the homologousrecombination vector, the altered portion of the MEMX gene is flanked atits 5′- and 3′-termini by additional nucleic acid of the MEMX gene toallow for homologous recombination to occur between the exogenous MEMXgene carried by the vector and an endogenous MEMX gene in an embryonicstem cell. The additional flanking MEMX nucleic acid is of sufficientlength for successful homologous recombination with the endogenous gene.Typically, several kilobases of flanking DNA (both at the 5′- and3′-termini) are included in the vector. See, e.g., Thomas, et al., 1987.Cell 51: 503 for a description of homologous recombination vectors. Thevector is ten introduced into an embryonic stem cell line (e.g., byelectroporation) and cells in which the introduced MEMX gene hashomologously-recombined with the endogenous MEMX gene are selected. See,e.g., Li, et al., 1992. Cell 69: 915.

[0286] The selected cells are then injected into a blastocyst of ananimal (e.g., a mouse) to form aggregation chimeras. See, e.g., Bradley,1987. In: TERATOCARCINOMAS AND EMBRYONIC STEM CELLS: A PRACTICALAPPROACH, Robertson, ed. IRL, Oxford, pp. 113-152. A chimeric embryo canthen be implanted into a suitable pseudopregnant female foster animaland the embryo brought to term. Progeny harboring thehomologously-recombined DNA in their germ cells can be used to breedanimals in which all cells of the animal contain thehomologously-recombined DNA by germline transmission of the transgene.Methods for constructing homologous recombination vectors and homologousrecombinant animals are described further in Bradley, 1991. Curr. Opin.Biotechnol. 2: 823-829; PCT International Publication Nos.: WO 90/11354;WO 91/01140; WO 92/0968; and WO 93/04169.

[0287] In another embodiment, transgenic non-humans animals can beproduced that contain selected systems that allow for regulatedexpression of the transgene. One example of such a system is thecre/loxP recombinase system of bacteriophage P1. For a description ofthe cre/loxP recombinase system, See, e.g., Lakso, et al., 1992. Proc.Natl. Acad. Sci. USA 89: 6232-6236. Another example of a recombinasesystem is the FLP recombinase system of Saccharomyces cerevisiae. See,O'Gorman, et al., 1991. Science 251:1351-1355. If a cre/loxP recombinasesystem is used to regulate expression of the transgene, animalscontaining transgenes encoding both the Cre recombinase and a selectedprotein are required. Such animals can be provided through theconstruction of “double” transgenic animals, e.g., by mating twotransgenic animals, one containing a transgene encoding a selectedprotein and the other containing a transgene encoding a recombinase.

[0288] Clones of the non-human transgenic animals described herein canalso be produced according to the methods described in Wilmut, et al.,1997. Nature 385: 810-813. In brief, a cell (e.g., a somatic cell) fromthe transgenic animal can be isolated and induced to exit the growthcycle and enter G₀ phase. The quiescent cell can then be fused, e.g.,through the use of electrical pulses, to an enucleated oocyte from ananimal of the same species from which the quiescent cell is isolated.The reconstructed oocyte is then cultured such that it develops tomorula or blastocyte and then transferred to pseudopregnant femalefoster animal. The offspring borne of this female foster animal will bea clone of the animal from which the cell (e.g., the somatic cell) isisolated.

[0289] Pharmaceutical Compositions

[0290] The MEMX nucleic acid molecules, MEMX proteins, and anti-MEMXantibodies (also referred to herein as “active compounds”) of theinvention, and derivatives, fragments, analogs and homologs thereof, canbe incorporated into pharmaceutical compositions suitable foradministration. Such compositions typically comprise the nucleic acidmolecule, protein, or antibody and a pharmaceutically acceptablecarrier. As used herein, “pharmaceutically acceptable carrier” isintended to include any and all solvents, dispersion media, coatings,antibacterial and antifungal agents, isotonic and absorption delayingagents, and the like, compatible with pharmaceutical administration.Suitable carriers are described in the most recent edition ofRemington's Pharmaceutical Sciences, a standard reference text in thefield, which is incorporated herein by reference. Preferred examples ofsuch carriers or diluents include, but are not limited to, water,saline, finger's solutions, dextrose solution, and 5% human serumalbumin. Liposomes and non-aqueous vehicles such as fixed oils may alsobe used. The use of such media and agents for pharmaceutically activesubstances is well known in the art. Except insofar as any conventionalmedia or agent is incompatible with the active compound, use thereof inthe compositions is contemplated. Supplementary active compounds canalso be incorporated into the compositions.

[0291] A pharmaceutical composition of the invention is formulated to becompatible with its intended route of administration. Examples of routesof administration include parenteral, e.g., intravenous, intradermal,subcutaneous, oral (e.g., inhalation), transdermal (i.e., topical),transmucosal, and rectal administration. Solutions or suspensions usedfor parenteral, intradermal, or subcutaneous application can include thefollowing components: a sterile diluent such as water for injection,saline solution, fixed oils, polyethylene glycols, glycerine, propyleneglycol or other synthetic solvents; antibacterial agents such as benzylalcohol or methyl parabens; antioxidants such as ascorbic acid or sodiumbisulfite; chelating agents such as ethylenediaminetetraacetic acid(EDTA); buffers such as acetates, citrates or phosphates, and agents forthe adjustment of tonicity such as sodium chloride or dextrose. The pHcan be adjusted with acids or bases, such as hydrochloric acid or sodiumhydroxide. The parenteral preparation can be enclosed in ampoules,disposable syringes or multiple dose vials made of glass or plastic.

[0292] Pharmaceutical compositions suitable for injectable use includesterile aqueous solutions (where water soluble) or dispersions andsterile powders for the extemporaneous preparation of sterile injectablesolutions or dispersion. For intravenous administration, suitablecarriers include physiological saline, bacteriostatic water, CremophorEL™ (BASF, Parsippany, N.J.) or phosphate buffered saline (PBS). In allcases, the composition must be sterile and should be fluid to the extentthat easy syringeability exists. It must be stable under the conditionsof manufacture and storage and must be preserved against thecontaminating action of microorganisms such as bacteria and fungi. Thecarrier can be a solvent or dispersion medium containing, for example,water, ethanol, polyol (for example, glycerol, propylene glycol, andliquid polyethylene glycol, and the like), and suitable mixturesthereof. The proper fluidity can be maintained, for example, by the useof a coating such as lecithin, by the maintenance of the requiredparticle size in the case of dispersion and by the use of surfactants.Prevention of the action of microorganisms can be achieved by variousantibacterial and antifungal agents, for example, parabens,chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In manycases, it will be preferable to include isotonic agents, for example,sugars, polyalcohols such as manitol, sorbitol, sodium chloride in thecomposition. Prolonged absorption of the injectable compositions can bebrought about by including in the composition an agent which delaysabsorption, for example, aluminum monostearate and gelatin.

[0293] Sterile injectable solutions can be prepared by incorporating theactive compound (e.g., an MEMX protein or anti-MEMX antibody) in therequired amount in an appropriate solvent with one or a combination ofingredients enumerated above, as required, followed by filteredsterilization. Generally, dispersions are prepared by incorporating theactive compound into a sterile vehicle that contains a basic dispersionmedium and the required other ingredients from those enumerated above.In the case of sterile powders for the preparation of sterile injectablesolutions, methods of preparation are vacuum drying and freeze-dryingthat yields a powder of the active ingredient plus any additionaldesired ingredient from a previously sterile-filtered solution thereof.

[0294] Oral compositions generally include an inert diluent or an ediblecarrier. They can be enclosed in gelatin capsules or compressed intotablets. For the purpose of oral therapeutic administration, the activecompound can be incorporated with excipients and used in the form oftablets, troches, or capsules. Oral compositions can also be preparedusing a fluid carrier for use as a mouthwash, wherein the compound inthe fluid carrier is applied orally and swished and expectorated orswallowed. Pharmaceutically compatible binding agents, and/or adjuvantmaterials can be included as part of the composition. The tablets,pills, capsules, troches and the like can contain any of the followingingredients, or compounds of a similar nature: a binder such asmicrocrystalline cellulose, gum tragacanth or gelatin; an excipient suchas starch or lactose, a disintegrating agent such as alginic acid,Primogel, or corn starch; a lubricant such as magnesium stearate orSterotes; a glidant such as colloidal silicon dioxide; a sweeteningagent such as sucrose or saccharin; or a flavoring agent such aspeppermint, methyl salicylate, or orange flavoring.

[0295] For administration by inhalation, the compounds are delivered inthe form of an aerosol spray from pressured container or dispenser whichcontains a suitable propellant, e.g., a gas such as carbon dioxide, or anebulizer.

[0296] Systemic administration can also be by transmucosal ortransdermal means. For transmucosal or transdermal administration,penetrants appropriate to the barrier to be permeated are used in theformulation. Such penetrants are generally known in the art, andinclude, for example, for transmucosal administration, detergents, bilesalts, and fusidic acid derivatives. Transmucosal administration can beaccomplished through the use of nasal sprays or suppositories. Fortransdermal administration, the active compounds are formulated intoointments, salves, gels, or creams as generally known in the art.

[0297] The compounds can also be prepared in the form of suppositories(e.g., with conventional suppository bases such as cocoa butter andother glycerides) or retention enemas for rectal delivery.

[0298] In one embodiment, the active compounds are prepared withcarriers that will protect the compound against rapid elimination fromthe body, such as a controlled release formulation, including implantsand microencapsulated delivery systems. Biodegradable, biocompatiblepolymers can be used, such as ethylene vinyl acetate, polyanhydrides,polyglycolic acid, collagen, polyorthoesters, and polylactic acid.Methods for preparation of such formulations will be apparent to thoseskilled in the art. The materials can also be obtained commercially fromAlza Corporation and Nova Pharmaceuticals, Inc. Liposomal suspensions(including liposomes targeted to infected cells with monoclonalantibodies to viral antigens) can also be used as pharmaceuticallyacceptable carriers. These can be prepared according to methods known tothose skilled in the art, for example, as described in U.S. Pat. No.4,522,811.

[0299] It is especially advantageous to formulate oral or parenteralcompositions in dosage unit form for ease of administration anduniformity of dosage. Dosage unit form as used herein refers tophysically discrete units suited as unitary dosages for the subject tobe treated; each unit containing a predetermined quantity of activecompound calculated to produce the desired therapeutic effect inassociation with the required pharmaceutical carrier. The specificationfor the dosage unit forms of the invention are dictated by and directlydependent on the unique characteristics of the active compound and theparticular therapeutic effect to be achieved, and the limitationsinherent in the art of compounding such an active compound for thetreatment of individuals.

[0300] The nucleic acid molecules of the invention can be inserted intovectors and used as gene therapy vectors. Gene therapy vectors can bedelivered to a subject by, for example, intravenous injection, localadministration (see, e.g., U.S. Pat. No. 5,328,470) or by stereotacticinjection (see, e.g., Chen, et al., 1994. Proc. Natl. Acad. Sci. USA 91:3054-3057). The pharmaceutical preparation of the gene therapy vectorcan include the gene therapy vector in an acceptable diluent, or cancomprise a slow release matrix in which the gene delivery vehicle isimbedded. Alternatively, where the complete gene delivery vector can beproduced intact from recombinant cells, e.g., retroviral vectors, thepharmaceutical preparation can include one or more cells that producethe gene delivery system.

[0301] The pharmaceutical compositions can be included in a container,pack, or dispenser together with instructions for administration.

[0302] Screening and Detection Methods

[0303] The isolated nucleic acid molecules of the invention can be usedto express MEMX protein (e.g., via a recombinant expression vector in ahost cell in gene therapy applications), to detect MEMX mRNA (e.g., in abiological sample) or a genetic lesion in an MEMX gene, and to modulateMEMX activity, as described further, below. In addition, the MEMXproteins can be used to screen drugs or compounds that modulate the MEMXprotein activity or expression as well as to treat disorderscharacterized by insufficient or excessive production of MEMX protein orproduction of MEMX protein forms that have decreased or aberrantactivity compared to MEMX wild-type protein. In addition, the anti-MEMXantibodies of the invention can be used to detect and isolate MEMXproteins and modulate MEMX activity.

[0304] The invention further pertains to novel agents identified by thescreening assays described herein and uses thereof for treatments asdescribed, supra.

[0305] I. Screening Assays

[0306] The invention provides a method (also referred to herein as a“screening assay”) for identifying modulators, i.e., candidate or testcompounds or agents (e.g., peptides, peptidomimetics, small molecules orother drugs) that bind to MEMX proteins or have a stimulatory orinhibitory effect on, e.g., MEMX protein expression or MEMX proteinactivity. The invention also includes compounds identified in thescreening assays described herein.

[0307] In one embodiment, the invention provides assays for screeningcandidate or test compounds which bind to or modulate the activity ofthe membrane-bound form of an MEMX protein or polypeptide orbiologically-active portion thereof. The test compounds of the inventioncan be obtained using any of the numerous approaches in combinatoriallibrary methods known in the art, including: biological libraries;spatially addressable parallel solid phase or solution phase libraries;synthetic library methods requiring deconvolution; the “one-beadone-compound” library method; and synthetic library methods usingaffinity chromatography selection. The biological library approach islimited to peptide libraries, while the other four approaches areapplicable to peptide, non-peptide oligomer or small molecule librariesof compounds. See, e.g., Lam, 1997. Anticancer Drug Design 12: 145.

[0308] A “small molecule” as used herein, is meant to refer to acomposition that has a molecular weight of less than about 5 Kdal andmost preferably less than about 4 Kdal. Small molecules can be, e.g.,nucleic acids, peptides, polypeptides, peptidomimetics, carbohydrates,lipids or other organic or inorganic molecules. Libraries of chemicaland/or biological mixtures, such as fungal, bacterial, or algalextracts, are known in the art and can be screened with any of theassays of the invention.

[0309] Examples of methods for the synthesis of molecular libraries canbe found in the art, for example in: DeWitt, et al., 1993. Proc. Natl.Acad. Sci. US.A. 90: 6909; Erb, et al., 1994. Proc. Natl. Acad. Sci.U.S.A. 91: 11422; Zuckermann, et al., 1994. J. Med Chem. 37: 2678; Cho,et al., 1993. Science 261: 1303; Carrell, et al., 1994. Angew. Chem.Int. Ed. Engl. 33: 2059; Carell, et al., 1994. Angew. Chem. Int. Ed.Engl. 33: 2061; Gallop, et al., 1994. J. Med. Chem. 37:1233.

[0310] Libraries of compounds may be presented in solution (e.g.,Houghten, 1992. Biotechniques 13: 412-421), or on beads (Lam, 1991.Nature 354: 82-84), on chips (Fodor, 1993. Nature 364: 555-556),bacteria (Ladner, U.S. Pat. No. 5,223,409), spores (Ladner, U.S. Pat.No. 5,233,409), plasmids (Cull, et al., 1992. Proc. Natl. Acad. Sci. USA89: 1865-1869) or on phage (Scott and Smith, 1990. Science 249: 386-390;Devlin, 1990. Science 249: 404-406; Cwirla, et al., 1990. Proc. Natl.Acad. Sci. U.S.A. 87: 6378-6382; Felici, 1991. J. Mol. Biol. 222:301-310; Ladner, U.S. Pat. No. 5,233,409.).

[0311] In one embodiment, an assay is a cell-based assay in which a cellwhich expresses a membrane-bound form of MEMX protein, or abiologically-active portion thereof, on the cell surface is contactedwith a test compound and the ability of the test compound to bind to anMEMX protein determined. The cell, for example, can of mammalian originor a yeast cell. Determining the ability of the test compound to bind tothe MEMX protein can be accomplished, for example, by coupling the testcompound with a radioisotope or enzymatic label such that binding of thetest compound to the MEMX protein or biologically-active portion thereofcan be determined by detecting the labeled compound in a complex. Forexample, test compounds can be labeled with ¹²⁵I, ³⁵S, ¹⁴C, or ³H,either directly or indirectly, and the radioisotope detected by directcounting of radioemission or by scintillation counting. Alternatively,test compounds can be enzymatically-labeled with, for example,horseradish peroxidase, alkaline phosphatase, or luciferase, and theenzymatic label detected by determination of conversion of anappropriate substrate to product. In one embodiment, the assay comprisescontacting a cell which expresses a membrane-bound form of MEMX protein,or a biologically-active portion thereof, on the cell surface with aknown compound which binds MEMX to form an assay mixture, contacting theassay mixture with a test compound, and determining the ability of thetest compound to interact with an MEMX protein, wherein determining theability of the test compound to interact with an MEMX protein comprisesdetermining the ability of the test compound to preferentially bind toMEMX protein or a biologically-active portion thereof as compared to theknown compound.

[0312] In another embodiment, an assay is a cell-based assay comprisingcontacting a cell expressing a membrane-bound form of MEMX protein, or abiologically-active portion thereof, on the cell surface with a testcompound and determining the ability of the test compound to modulate(e.g., stimulate or inhibit) the activity of the MEMX protein orbiologically-active portion thereof. Determining the ability of the testcompound to modulate the activity of MEMX or a biologically-activeportion thereof can be accomplished, for example, by determining theability of the MEMX protein to bind to or interact with an MEMX targetmolecule. As used herein, a “target molecule” is a molecule with whichan MEMX protein binds or interacts in nature, for example, a molecule onthe surface of a cell which expresses an MEMX interacting protein, amolecule on the surface of a second cell, a molecule in theextracellular milieu, a molecule associated with the internal surface ofa cell membrane or a cytoplasmic molecule. An MEMX target molecule canbe a non-MEMX molecule or an MEMX protein or polypeptide of theinvention . In one embodiment, an MEMX target molecule is a component ofa signal transduction pathway that facilitates transduction of anextracellular signal (e.g. a signal generated by binding of a compoundto a membrane-bound MEMX molecule) through the cell membrane and intothe cell. The target, for example, can be a second intercellular proteinthat has catalytic activity or a protein that facilitates theassociation of downstream signaling molecules with MEMX.

[0313] Determining the ability of the MEMX protein to bind to orinteract with an MEMX target molecule can be accomplished by one of themethods described above for determining direct binding. In oneembodiment, determining the ability of the MEMX protein to bind to orinteract with an MEMX target molecule can be accomplished by determiningthe activity of the target molecule. For example, the activity of thetarget molecule can be determined by detecting induction of a cellularsecond messenger of the target (i.e. intracellular Ca²⁺, diacylglycerol,IP₃, etc.), detecting catalytic/enzymatic activity of the target anappropriate substrate, detecting the induction of a reporter gene(comprising an MEMX-responsive regulatory element operatively linked toa nucleic acid encoding a detectable marker, e.g., luciferase), ordetecting a cellular response, for example, cell survival, cellulardifferentiation, or cell proliferation.

[0314] In yet another embodiment, an assay of the invention is acell-free assay comprising contacting an MEMX protein orbiologically-active portion thereof with a test compound and determiningthe ability of the test compound to bind to the MEMX protein orbiologically-active portion thereof. Binding of the test compound to theMEMX protein can be determined either directly or indirectly asdescribed above. In one such embodiment, the assay comprises contactingthe MEMX protein or biologically-active portion thereof with a knowncompound which binds MEMX to form an assay mixture, contacting the assaymixture with a test compound, and determining the ability of the testcompound to interact with an MEMX protein, wherein determining theability of the test compound to interact with an MEMX protein comprisesdetermining the ability of the test compound to preferentially bind toMEMX or biologically-active portion thereof as compared to the knowncompound.

[0315] In still another embodiment, an assay is a cell-free assaycomprising contacting MEMX protein or biologically-active portionthereof with a test compound and determining the ability of the testcompound to modulate (e.g. stimulate or inhibit) the activity of theMEMX protein or biologically-active portion thereof. Determining theability of the test compound to modulate the activity of MEMX can beaccomplished, for example, by determining the ability of the MEMXprotein to bind to an MEMX target molecule by one of the methodsdescribed above for determining direct binding. In an alternativeembodiment, determining the ability of the test compound to modulate theactivity of MEMX protein can be accomplished by determining the abilityof the MEMX protein further modulate an MEMX target molecule. Forexample, the catalytic/enzymatic activity of the target molecule on anappropriate substrate can be determined as described, supra.

[0316] In yet another embodiment, the cell-free assay comprisescontacting the MEMX protein or biologically-active portion thereof witha known compound which binds MEMX protein to form an assay mixture,contacting the assay mixture with a test compound, and determining theability of the test compound to interact with an MEMX protein, whereindetermining the ability of the test compound to interact with an MEMXprotein comprises determining the ability of the MEMX protein topreferentially bind to or modulate the activity of an MEMX targetmolecule.

[0317] The cell-free assays of the invention are amenable to use of boththe soluble form or the membrane-bound form of MEMX protein. In the caseof cell-free assays comprising the membrane-bound form of MEMX protein,it may be desirable to utilize a solubilizing agent such that themembrane-bound form of MEMX protein is maintained in solution. Examplesof such solubilizing agents include non-ionic detergents such asn-octylglucoside, n-dodecylglucoside, n-dodecylmaltoside,octanoyl-N-methylglucamide, decanoyl-N-methylglucamide, Triton® X-100,Triton® X-114, Thesit®, Isotridecypoly(ethylene glycol ether)_(n),N-dodecyl-N,N-dimethyl-3-ammonio-1-propane sulfonate,3-(3-cholamidopropyl) dimethylamminiol-1-propane sulfonate (CHAPS), or3-(3-cholamidopropyl)dimethylamminiol-2-hydroxy-1-propane sulfonate(CHAPSO).

[0318] In more than one embodiment of the above assay methods of theinvention, it may be desirable to immobilize either MEMX protein or itstarget molecule to facilitate separation of complexed from uncomplexedforms of one or both of the proteins, as well as to accommodateautomation of the assay. Binding of a test compound to MEMX protein, orinteraction of MEMX protein with a target molecule in the presence andabsence of a candidate compound, can be accomplished in any vesselsuitable for containing the reactants. Examples of such vessels includemicrotiter plates, test tubes, and micro-centrifuge tubes. In oneembodiment, a fusion protein can be provided that adds a domain thatallows one or both of the proteins to be bound to a matrix. For example,GST-MEMX fusion proteins or GST-target fusion proteins can be adsorbedonto glutathione sepharose beads (Sigma Chemical, St. Louis, Mo.) orglutathione derivatized microtiter plates, that are then combined withthe test compound or the test compound and either the non-adsorbedtarget protein or MEMX protein, and the mixture is incubated underconditions conducive to complex formation (e.g., at physiologicalconditions for salt and pH). Following incubation, the beads ormicrotiter plate wells are washed to remove any unbound components, thematrix immobilized in the case of beads, complex determined eitherdirectly or indirectly, for example, as described, supra. Alternatively,the complexes can be dissociated from the matrix, and the level of MEMXprotein binding or activity determined using standard techniques.

[0319] Other techniques for immobilizing proteins on matrices can alsobe used in the screening assays of the invention. For example, eitherthe MEMX protein or its target molecule can be immobilized utilizingconjugation of biotin and streptavidin. Biotinylated MEMX protein ortarget molecules can be prepared from biotin-NHS (N-hydroxy-succinimide)using techniques well-known within the art (e.g., biotinylation kit,Pierce Chemicals, Rockford, Ill.), and immobilized in the wells ofstreptavidin-coated 96 well plates (Pierce Chemical). Alternatively,antibodies reactive with MEMX protein or target molecules, but which donot interfere with binding of the MEMX protein to its target molecule,can be derivatized to the wells of the plate, and unbound target or MEMXprotein trapped in the wells by antibody conjugation. Methods fordetecting such complexes, in addition to those described above for theGST-immobilized complexes, include immunodetection of complexes usingantibodies reactive with the MEMX protein or target molecule, as well asenzyme-linked assays that rely on detecting an enzymatic activityassociated with the MEMX protein or target molecule.

[0320] In another embodiment, modulators of MEMX protein expression areidentified in a method wherein a cell is contacted with a candidatecompound and the expression of MEMX mRNA or protein in the cell isdetermined. The level of expression of MEMX mRNA or protein in thepresence of the candidate compound is compared to the level ofexpression of MEMX mRNA or protein in the absence of the candidatecompound. The candidate compound can then be identified as a modulatorof MEMX mRNA or protein expression based upon this comparison. Forexample, when expression of MEMX mRNA or protein is greater (i.e.,statistically significantly greater) in the presence of the candidatecompound than in its absence, the candidate compound is identified as astimulator of MEMX mRNA or protein expression. Alternatively, whenexpression of MEMX mRNA or protein is less (statistically significantlyless) in the presence of the candidate compound than in its absence, thecandidate compound is identified as an inhibitor of MEMX mRNA or proteinexpression. The level of MEMX mRNA or protein expression in the cellscan be determined by methods described herein for detecting MEMX mRNA orprotein.

[0321] In yet another aspect of the invention, the MEMX proteins can beused as “bait proteins” in a two-hybrid assay or three hybrid assay(see, e.g., U.S. Pat. No. 5,283,317; Zervos, et al., 1993. Cell 72:223-232; Madura, et al., 1993. J. Biol. Chem. 268: 12046-12054; Bartel,et al., 1993. Biotechniques 14: 920-924; Iwabuchi, et al., 1993.Oncogene 8: 1693-1696; and Brent WO 94/10300), to identify otherproteins that bind to or interact with MEMX (“MEMX-binding proteins” or“MEMX-bp”) and modulate MEMX activity. Such MEMX-binding proteins arealso likely to be involved in the propagation of signals by the MEMXproteins as, for example, upstream or downstream elements of the MEMXpathway.

[0322] The two-hybrid system is based on the modular nature of mosttranscription factors, which consist of separable DNA-binding andactivation domains. Briefly, the assay utilizes two different DNAconstructs. In one construct, the gene that codes for MEMX is fused to agene encoding the DNA binding domain of a known transcription factor(e.g., GAL-4). In the other construct, a DNA sequence, from a library ofDNA sequences, that encodes an unidentified protein (“prey” or “sample”)is fused to a gene that codes for the activation domain of the knowntranscription factor. If the “bait” and the “prey” proteins are able tointeract, in vivo, forming an MEMX-dependent complex, the DNA-bindingand activation domains of the transcription factor are brought intoclose proximity. This proximity allows transcription of a reporter gene(e.g., LacZ) that is operably linked to a transcriptional regulatorysite responsive to the transcription factor. Expression of the reportergene can be detected and cell colonies containing the functionaltranscription factor can be isolated and used to obtain the cloned genethat encodes the protein which interacts with MEMX.

[0323] The invention further pertains to novel agents identified by theaforementioned screening assays and uses thereof for treatments asdescribed herein.

[0324] II. Detection Assays

[0325] Portions or fragments of the cDNA sequences identified herein(and the corresponding complete gene sequences) can be used in numerousways as polynucleotide reagents. By way of example, and not oflimitation, these sequences can be used to: (i) map their respectivegenes on a chromosome; and, thus, locate gene regions associated withgenetic disease; (ii) identify an individual from a minute biologicalsample (tissue typing); and (iii) aid in forensic identification of abiological sample. Some of these applications are described in thesubsections, below.

[0326] Chromosome Mapping

[0327] Once the sequence (or a portion of the sequence) of a gene hasbeen isolated, this sequence can be used to map the location of the geneon a chromosome. This process is called chromosome mapping. Accordingly,portions or fragments of the MEMX sequences, SEQ ID NO:1, 3, 5, 7, 9,11, 13, or 15, or fragments or derivatives thereof, can be used to mapthe location of the MEMX genes, respectively, on a chromosome. Themapping of the MEMX sequences to chromosomes is an important first stepin correlating these sequences with genes associated with disease.

[0328] Briefly, MEMX genes can be mapped to chromosomes by preparing PCRprimers (preferably 15-25 bp in length) from the MEMX sequences.Computer analysis of the MEMX, sequences can be used to rapidly selectprimers that do not span more than one exon in the genomic DNA, thuscomplicating the amplification process. These primers can then be usedfor PCR screening of somatic cell hybrids containing individual humanchromosomes. Only those hybrids containing the human gene correspondingto the MEMX sequences will yield an amplified fragment.

[0329] Somatic cell hybrids are prepared by fusing somatic cells fromdifferent mammals (e.g., human and mouse cells). As hybrids of human andmouse cells grow and divide, they gradually lose human chromosomes inrandom order, but retain the mouse chromosomes. By using media in whichmouse cells cannot grow, because they lack a particular enzyme, but inwhich human cells can, the one human chromosome that contains the geneencoding the needed enzyme will be retained. By using various media,panels of hybrid cell lines can be established. Each cell line in apanel contains either a single human chromosome or a small number ofhuman chromosomes, and a full set of mouse chromosomes, allowing easymapping of individual genes to specific human chromosomes. See, e.g.,D'Eustachio, et al., 1983. Science 220: 919-924. Somatic cell hybridscontaining only fragments of human chromosomes can also be produced byusing human chromosomes with translocations and deletions.

[0330] PCR mapping of somatic cell hybrids is a rapid procedure forassigning a particular sequence to a particular chromosome. Three ormore sequences can be assigned per day using a single thermal cycler.Using the MEMX sequences to design oligonucleotide primers,sub-localization can be achieved with panels of fragments from specificchromosomes.

[0331] Fluorescence in situ hybridization (FISH) of a DNA sequence to ametaphase chromosomal spread can further be used to provide a precisechromosomal location in one step. Chromosome spreads can be made usingcells whose division has been blocked in metaphase by a chemical likecolcemid that disrupts the mitotic spindle. The chromosomes can betreated briefly with trypsin, and then stained with Giemsa. A pattern oflight and dark bands develops on each chromosome, so that thechromosomes can be identified individually. The FISH technique can beused with a DNA sequence as short as 500 or 600 bases. However, cloneslarger than 1,000 bases have a higher likelihood of binding to a uniquechromosomal location with sufficient signal intensity for simpledetection. Preferably 1,000 bases, and more preferably 2,000 bases, willsuffice to get good results at a reasonable amount of time. For a reviewof this technique, see, Verma, et al., HUMAN CHROMOSOMES: A MANUAL OFBASIC TECHNIQUES (Pergamon Press, New York 1988).

[0332] Reagents for chromosome mapping can be used individually to marka single chromosome or a single site on that chromosome, or panels ofreagents can be used for marking multiple sites and/or multiplechromosomes. Reagents corresponding to non-coding regions of the genesactually are preferred for mapping purposes. Coding sequences are morelikely to be conserved within gene families, thus increasing the chanceof cross hybridizations during chromosomal mapping.

[0333] Once a sequence has been mapped to a precise chromosomallocation, the physical position of the sequence on the chromosome can becorrelated with genetic map data. Such data are found, e.g., inMcKusick, MENDELIAN INHERITANCE IN MAN, available on-line through JohnsHopkins University Welch Medical Library). The relationship betweengenes and disease, mapped to the same chromosomal region, can then beidentified through linkage analysis (co-inheritance of physicallyadjacent genes), described in, e.g., Egeland, et al., 1987. Nature, 325:783-787.

[0334] Moreover, differences in the DNA sequences between individualsaffected and unaffected with a disease associated with the MEMX gene,can be determined. If a mutation is observed in some or all of theaffected individuals but not in any unaffected individuals, then themutation is likely to be the causative agent of the particular disease.Comparison of affected and unaffected individuals generally involvesfirst looking for structural alterations in the chromosomes, such asdeletions or translocations that are visible from chromosome spreads ordetectable using PCR based on that DNA sequence. Ultimately, completesequencing of genes from several individuals can be performed to confirmthe presence of a mutation and to distinguish mutations frompolymorphisms.

[0335] Tissue Typing

[0336] The MEMX sequences of the invention can also be used to identifyindividuals from minute biological samples. In this technique, anindividual's genomic DNA is digested with one or more restrictionenzymes, and probed on a Southern blot to yield unique bands foridentification. The sequences of the invention are useful as additionalDNA markers for Restriction Fragment Length Polymorphisms (RFLP)described in U.S. Pat. No. 5,272,057.

[0337] Furthermore, the sequences of the invention can be used toprovide an alternative technique that determines the actual base-by-baseDNA sequence of selected portions of an individual's genome. Thus, theMEMX sequences described herein can be used to prepare two PCR primersfrom the 5′- and 3′-termini of the sequences. These primers can then beused to amplify an individual's DNA and subsequently sequence it.

[0338] Panels of corresponding DNA sequences from individuals, preparedin this manner, can provide unique individual identifications, as eachindividual will have a unique set of such DNA sequences due to allelicdifferences. The sequences of the invention can be used to obtain suchidentification sequences from individuals and from tissue. The MEMXsequences of the invention uniquely represent portions of the humangenome. Allelic variation occurs to some degree in the coding regions ofthese sequences, and to a greater degree in the non-coding regions. Itis estimated that allelic variation between individual humans occurswith a frequency of about once per each 500 bases. Much of the allelicvariation is due to single nucleotide polymorphisms (SNPs), whichinclude RFLPs.

[0339] Each of the sequences described herein can, to some degree, beused as a standard against which DNA from an individual can be comparedfor identification purposes. Because greater numbers of polymorphismsoccur in the non-coding regions, fewer sequences are necessary todifferentiate individuals. The non-coding sequences can comfortablyprovide positive individual identification with a panel of perhaps 10 to1,000 primers that each yield a non-coding amplified sequence of 100bases. If predicted coding sequences, such as those in SEQ ID NO:1, 3,5, 7, 9, 11, 13, or 15, are used, a more appropriate number of primersfor positive individual identification would be 500-2,000.

[0340] Predictive Medicine

[0341] The invention also pertains to the field of predictive medicinein which diagnostic assays, prognostic assays, pharmacogenomics, andmonitoring clinical trials are used for prognostic (predictive) purposesto thereby treat an individual prophylactically. Accordingly, one aspectof the invention relates to diagnostic assays for determining MEMXprotein and/or nucleic acid expression as well as MEMX activity, in thecontext of a biological sample (e.g., blood, serum, cells, tissue) tothereby determine whether an individual is afflicted with a disease ordisorder, or is at risk of developing a disorder, associated withaberrant MEMX expression or activity. The invention also provides forprognostic (or predictive) assays for determining whether an individualis at risk of developing a disorder associated with MEMX protein,nucleic acid expression or activity. For example, mutations in an MEMXgene can be assayed in a biological sample. Such assays can be used forprognostic or predictive purpose to thereby prophylactically treat anindividual prior to the onset of a disorder characterized by orassociated with MEMX protein, nucleic acid expression, or biologicalactivity.

[0342] Another aspect of the invention provides methods for determiningMEMX protein, nucleic acid expression or activity in an individual tothereby select appropriate therapeutic or prophylactic agents for thatindividual (referred to herein as “pharmacogenomics”). Pharmacogenomicsallows for the selection of agents (e.g., drugs) for therapeutic orprophylactic treatment of an individual based on the genotype of theindividual (e.g., the genotype of the individual examined to determinethe ability of the individual to respond to a particular agent.).

[0343] Yet another aspect of the invention pertains to monitoring theinfluence of agents (e.g., drugs, compounds) on the expression oractivity of MEMX in clinical trials.

[0344] These and other agents are described in further detail in thefollowing sections.

[0345] I. Diagnostic Assays

[0346] An exemplary method for detecting the presence or absence of MEMXin a biological sample involves obtaining a biological sample from atest subject and contacting the biological sample with a compound or anagent capable of detecting MEMX protein or nucleic acid (e.g., mRNA,genomic DNA) that encodes MEMX protein such that the presence of MEMX isdetected in the biological sample. An agent for detecting MEMX mRNA orgenomic DNA is a labeled nucleic acid probe capable of hybridizing toMEMX mRNA or genomic DNA. The nucleic acid probe can be, for example, afull-length MEMX nucleic acid, such as the nucleic acid of SEQ ID NO:1,3, 5 7, 9, 11, 13, or 15, or a portion thereof, such as anoligonucleotide of at least 15, 30, 50, 100, 250 or 500 nucleotides inlength and sufficient to specifically hybridize under stringentconditions to MEMX mRNA or genomic DNA. Other suitable probes for use inthe diagnostic assays of the invention are described herein.

[0347] One agent for detecting MEMX protein is an antibody capable ofbinding to MEMX protein, preferably an antibody with a detectable label.Antibodies directed against a protein of the invention may be used inmethods known within the art relating to the localization and/orquantitation of the protein (e.g., for use in measuring levels of theprotein within appropriate physiological samples, for use in diagnosticmethods, for use in imaging the protein, and the like). In a givenembodiment, antibodies against the proteins, or derivatives, fragments,analogs or homologs thereof, that contain the antigen binding domain,are utilized as pharmacologically-active compounds.

[0348] An antibody specific for a protein of the invention can be usedto isolate the protein by standard techniques, such as immunoaffinitychromatography or immunoprecipitation. Such an antibody can facilitatethe purification of the natural protein antigen from cells and ofrecombinantly produced antigen expressed in host cells. Moreover, suchan antibody can be used to detect the antigenic protein (e.g., in acellular lysate or cell supernatant) in order to evaluate the abundanceand pattern of expression of the antigenic protein. Antibodies directedagainst the protein can be used diagnostically to monitor protein levelsin tissue as part of a clinical testing procedure, e.g., to, forexample, determine the efficacy of a given treatment regimen. Detectioncan be facilitated by coupling (i.e., physically linking) the antibodyto a detectable substance. Examples of detectable substances includevarious enzymes, prosthetic groups, fluorescent materials, luminescentmaterials, bioluminescent materials, and radioactive materials. Examplesof suitable enzymes include horseradish peroxidase, alkalinephosphatase, -g alactosidase, or acetylcholinesterase; examples ofsuitable prosthetic group complexes include streptavidin/biotin andavidin/biotin; examples of suitable fluorescent materials includeumbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine,dichlorotriazinylamine fluorescein, dansyl chloride or phycoerythrin; anexample of a luminescent material includes luminol; examples ofbioluminescent materials include luciferase, luciferin, and aequorin,and examples of suitable radioactive material include ¹²⁵I, ¹³¹I, ³⁵S or³H.

[0349] Antibodies can be polyclonal, or more preferably, monoclonal. Anintact antibody, or a fragment thereof (e.g., Fab or F(ab′)₂) can beused. The term “labeled”, with regard to the probe or antibody, isintended to encompass direct labeling of the probe or antibody bycoupling (i.e., physically linking) a detectable substance to the probeor antibody, as well as indirect labeling of the probe or antibody byreactivity with another reagent that is directly labeled. Examples ofindirect labeling include detection of a primary antibody using afluorescently-labeled secondary antibody and end-labeling of a DNA probewith biotin such that it can be detected with fluorescently-labeledstreptavidin. The term “biological sample” is intended to includetissues, cells and biological fluids isolated from a subject, as well astissues, cells and fluids present within a subject. That is, thedetection method of the invention can be used to detect MEMX mRNA,protein, or genomic DNA in a biological sample in vitro as well as invivo. For example, in vitro techniques for detection of MEMX mRNAinclude Northern hybridizations and in situ hybridizations. In vitrotechniques for detection of MEMX protein include enzyme linkedimmunosorbent assays (ELISAs), Western blots, immunoprecipitations, andimmunofluorescence. In vitro techniques for detection of MEMX genomicDNA include Southern hybridizations. Furthermore, in vivo techniques fordetection of MEMX protein include introducing into a subject a labeledanti-MEMX antibody. For example, the antibody can be labeled with aradioactive marker whose presence and location in a subject can bedetected by standard imaging techniques.

[0350] In one embodiment, the biological sample contains proteinmolecules from the test subject. Alternatively, the biological samplecan contain mRNA molecules from the test subject or genomic DNAmolecules from the test subject. A preferred biological sample is aperipheral blood leukocyte sample isolated by conventional means from asubject.

[0351] In one embodiment, the methods further involve obtaining acontrol biological sample from a control subject, contacting the controlsample with a compound or agent capable of detecting MEMX protein, mRNA,or genomic DNA, such that the presence of MEMX protein, mRNA or genomicDNA is detected in the biological sample, and comparing the presence ofMEMX protein, MRNA or genomic DNA in the control sample with thepresence of MEMX protein, mRNA or genomic DNA in the test sample.

[0352] The invention also encompasses kits for detecting the presence ofMEMX in a biological sample. For example, the kit can comprise: alabeled compound or agent capable of detecting MEMX protein or MRNA in abiological sample; means for determining the amount of MEMX in thesample; and means for comparing the amount of MEMX in the sample with astandard. The compound or agent can be packaged in a suitable container.The kit can further comprise instructions for using the kit to detectMEMX protein or nucleic acid.

[0353] II. Prognostic Assays

[0354] The diagnostic methods described herein can furthermore beutilized to identify subjects having or at risk of developing a diseaseor disorder associated with aberrant MEMX expression or activity. Suchdisorders for MEMI include immunological conditions, viral infections,neurological disorders, Alzheimer's or Parkinson's Diseases, cancer(e.g., breast or neuroblastoma), nephrology, and female reproductivehealth. Such disorders for MEM4 include those involving the lung and/orbrain (e.g., schizophrenia, or neuronal damage following head injury).Disorders for MEM5 include heart and other muscular disorders (e.g.,arrhythmial), clotting deficiencies, and cobalamine deficiencies (e.g.,pernicious anemia). Such disorders for MEM6 include those originating indysregulation of glycogen metabolism (e.g., diabetes). Such disordersfor MEM7 and MEM8 include vision-related disorders (e.g.,keratomalacia), cancer, and other neoplastic pathologies.

[0355] For example, the assays described herein, such as the precedingdiagnostic assays or the following assays, can be utilized to identify asubject having or at risk of developing a disorder associated with MEMXprotein, nucleic acid expression or activity. Alternatively, theprognostic assays can be utilized to identify a subject having or atrisk for developing a disease or disorder. Thus, the invention providesa method for identifying a disease or disorder associated with aberrantMEMX expression or activity in which a test sample is obtained from asubject and MEMX protein or nucleic acid (e.g., mRNA, genomic DNA) isdetected, wherein the presence of MEMX protein or nucleic acid isdiagnostic for a subject having or at risk of developing a disease ordisorder associated with aberrant MEMX expression or activity. As usedherein, a “test sample” refers to a biological sample obtained from asubject of interest. For example, a test sample can be a biologicalfluid (e.g., serum), cell sample, or tissue.

[0356] Furthermore, the prognostic assays described herein can be usedto determine whether a subject can be administered an agent (e.g., anagonist, antagonist, peptidomimetic, protein, peptide, nucleic acid,small molecule, or other drug candidate) to treat a disease or disorderassociated with aberrant MEMX expression or activity. For example, suchmethods can be used to determine whether a subject can be effectivelytreated with an agent for a disorder. Thus, the invention providesmethods for determining whether a subject can be effectively treatedwith an agent for a disorder associated with aberrant MEMX expression oractivity in which a test sample is obtained and MEMX protein or nucleicacid is detected (e.g., wherein the presence of MEMX protein or nucleicacid is diagnostic for a subject that can be administered the agent totreat a disorder associated with aberrant MEMX expression or activity).

[0357] The methods of the invention can also be used to detect geneticlesions in an MEMX gene, thereby determining if a subject with thelesioned gene is at risk for a disorder characterized by aberrant cellproliferation and/or differentiation. In various embodiments, themethods include detecting, in a sample of cells from the subject, thepresence or absence of a genetic lesion characterized by at least one ofan alteration affecting the integrity of a gene encoding anMEMX-protein, or the misexpression of the MEMX gene. For example, suchgenetic lesions can be detected by ascertaining the existence of atleast one of: (i) a deletion of one or more nucleotides from an MEMXgene; (ii) an addition of one or more nucleotides to an MEMX gene; (iii)a substitution of one or more nucleotides of an MEMX gene, (iv) achromosomal rearrangement of an MEMX gene; (v) an alteration in thelevel of a messenger RNA transcript of an MEMX gene, (vi) aberrantmodification of an MEMX gene, such as of the methylation pattern of thegenomic DNA, (vii) the presence of a non-wild-type splicing pattern of amessenger RNA transcript of an MEMX gene, (viii) a non-wild-type levelof an MEMX protein, (ix) allelic loss of an MEMX gene, and (x)inappropriate post-translational modification of an MEMX protein. Asdescribed herein, there are a large number of assay techniques known inthe art which can be used for detecting lesions in an MEMX gene. Apreferred biological sample is a peripheral blood leukocyte sampleisolated by conventional means from a subject. However, any biologicalsample containing nucleated cells may be used, including, for example,buccal mucosal cells.

[0358] In certain embodiments, detection of the lesion involves the useof a probe/primer in a polymerase chain reaction (PCR) (see, e.g., U.S.Pat. Nos. 4,683,195 and 4,683,202), such as anchor PCR or RACE PCR, or,alternatively, in a ligation chain reaction (LCR) (see, e.g., Landegran,et al., 1988. Science 241: 1077-1080; and Nakazawa, et al., 1994. Proc.Natl. Acad. Sci. USA 91: 360-364), the latter of which can beparticularly useful for detecting point mutations in the MEMX-gene (see,Abravaya, et al., 1995. Nucl. Acids Res. 23: 675-682). This method caninclude the steps of collecting a sample of cells from a patient,isolating nucleic acid (e.g., genomic, mRNA or both) from the cells ofthe sample, contacting the nucleic acid sample with one or more primersthat specifically hybridize to an MEMX gene under conditions such thathybridization and amplification of the MEMX gene (if present) occurs,and detecting the presence or absence of an amplification product, ordetecting the size of the amplification product and comparing the lengthto a control sample. It is anticipated that PCR and/or LCR may bedesirable to use as a preliminary amplification step in conjunction withany of the techniques used for detecting mutations described herein.

[0359] Alternative amplification methods include: self sustainedsequence replication (see, Guatelli, et al., 1990. Proc. Natl. Acad.Sci. USA 87: 1874-1878), transcriptional amplification system (see,Kwoh, et al., 1989. Proc. Natl. Acad. Sci. USA 86: 1173-1177); QβReplicase (see, Lizardi, et al, 1988. BioTechnology 6: 1197), or anyother nucleic acid amplification method, followed by the detection ofthe amplified molecules using techniques well known to those of skill inthe art. These detection schemes are especially useful for the detectionof nucleic acid molecules if such molecules are present in very lownumbers.

[0360] In an alternative embodiment, mutations in an MEMX gene from asample cell can be identified by alterations in restriction enzymecleavage patterns. For example, sample and control DNA is isolated,amplified (optionally), digested with one or more restrictionendonucleases, and fragment length sizes are determined by gelelectrophoresis and compared. Differences in fragment length sizesbetween sample and control DNA indicates mutations in the sample DNA.Moreover, the use of sequence specific ribozymes (see, e.g., U.S. Pat.No. 5,493,531) can be used to score for the presence of specificmutations by development or loss of a ribozyme cleavage site.

[0361] In other embodiments, genetic mutations in MEMX can be identifiedby hybridizing a sample and control nucleic acids, e.g., DNA or RNA, tohigh-density arrays containing hundreds or thousands of oligonucleotidesprobes. See, e.g., Cronin, et al., 1996. Human Mutation 7: 244-255;Kozal, et al., 1996. Nat. Med. 2: 753-759. For example, geneticmutations in MEMX can be identified in two dimensional arrays containinglight-generated DNA probes as described in Cronin, et al., supra.Briefly, a first hybridization array of probes can be used to scanthrough long stretches of DNA in a sample and control to identify basechanges between the sequences by making linear arrays of sequentialoverlapping probes. This step allows the identification of pointmutations. This is followed by a second hybridization array that allowsthe characterization of specific mutations by using smaller, specializedprobe arrays complementary to all variants or mutations detected. Eachmutation array is composed of parallel probe sets, one complementary tothe wild-type gene and the other complementary to the mutant gene.

[0362] In yet another embodiment, any of a variety of sequencingreactions known in the art can be used to directly sequence the MEMXgene and detect mutations by comparing the sequence of the sample MEMXwith the corresponding wild-type (control) sequence. Examples ofsequencing reactions include those based on techniques developed byMaxim and Gilbert, 1977. Proc. Natl. Acad. Sci. USA 74: 560 or Sanger,1977. Proc. Natl. Acad. Sci. USA 74: 5463. It is also contemplated thatany of a variety of automated sequencing procedures can be utilized whenperforming the diagnostic assays (see, e.g., Naeve, et al., 1995.Biotechniques 19: 448), including sequencing by mass spectrometry (see,e.g., PCT International Publication No. WO 94/16101; Cohen, et al.,1996. Adv. Chromatography 36: 127-162; and Griffin, et al., 1993. Appl.Biochem. Biotechnol. 38: 147-159).

[0363] Other methods for detecting mutations in the MEMX gene includemethods in which protection from cleavage agents is used to detectmismatched bases in RNA/RNA or RNA/DNA heteroduplexes. See, e.g., Myers,et al., 1985. Science 230: 1242. In general, the art technique of“mismatch cleavage” starts by providing heteroduplexes of formed byhybridizing (labeled) RNA or DNA containing the wild-type MEMX sequencewith potentially mutant RNA or DNA obtained from a tissue sample. Thedouble-stranded duplexes are treated with an agent that cleavessingle-stranded regions of the duplex such as which will exist due tobasepair mismatches between the control and sample strands. Forinstance, RNA/DNA duplexes can be treated with RNase and DNA/DNA hybridstreated with SI nuclease to enzymatically digesting the mismatchedregions. In other embodiments, either DNA/DNA or RNA/DNA duplexes can betreated with hydroxylamine or osmium tetroxide and with piperidine inorder to digest mismatched regions. After digestion of the mismatchedregions, the resulting material is then separated by size on denaturingpolyacrylamide gels to determine the site of mutation. See, e.g.,Cotton, et al., 1988. Proc. Natl. Acad. Sci. USA 85: 4397; Saleeba, etal., 1992. Methods Enzymol. 217: 286-295. In an embodiment, the controlDNA or RNA can be labeled for detection.

[0364] In still another embodiment, the mismatch cleavage reactionemploys one or more proteins that recognize mismatched base pairs indouble-stranded DNA (so called “DNA mismatch repair” enzymes) in definedsystems for detecting and mapping point mutations in MEMX cDNAs obtainedfrom samples of cells. For example, the mutY enzyme of E. coli cleaves Aat G/A mismatches and the thymidine DNA glycosylase from HeLa cellscleaves T at G/T mismatches. See, e.g., Hsu, et al., 1994.Carcinogenesis 15: 1657-1662. According to an exemplary embodiment, aprobe based on an MEMX sequence, e.g., a wild-type MEMX sequence, ishybridized to a cDNA or other DNA product from a test cell(s). Theduplex is treated with a DNA mismatch repair enzyme, and the cleavageproducts, if any, can be detected from electrophoresis protocols or thelike. See, e.g., U.S. Pat. No. 5,459,039.

[0365] In other embodiments, alterations in electrophoretic mobilitywill be used to identify mutations in MEMX genes. For example, singlestrand conformation polymorphism (SSCP) may be used to detectdifferences in electrophoretic mobility between mutant and wild typenucleic acids. See, e.g., Orita, et al., 1989. Proc. Natl. Acad. Sci.USA: 86: 2766; Cotton, 1993. Mutat. Res. 285: 125-144; Hayashi, 1992.Genet. Anal. Tech. AppL. 9: 73-79. Single-stranded DNA fragments ofsample and control MEMX nucleic acids will be denatured and allowed torenature. The secondary structure of single-stranded nucleic acidsvaries according to sequence, the resulting alteration inelectrophoretic mobility enables the detection of even a single basechange. The DNA fragments may be labeled or detected with labeledprobes. The sensitivity of the assay may be enhanced by using RNA(rather than DNA), in which the secondary structure is more sensitive toa change in sequence. In one embodiment, the subject method utilizesheteroduplex analysis to separate double stranded heteroduplex moleculeson the basis of changes in electrophoretic mobility. See, e.g., Keen, etal., 1991. Trends Genet. 7: 5.

[0366] In yet another embodiment, the movement of mutant or wild-typefragments in polyacrylamide gels containing a gradient of denaturant isassayed using denaturing gradient gel electrophoresis (DGGE). See, e.g.,Myers, et al., 1985. Nature 313: 495. When DGGE is used as the method ofanalysis, DNA will be modified to insure that it does not completelydenature, for example by adding a GC clamp of approximately 40 bp ofhigh-melting GC-rich DNA by PCR. In a further embodiment, a temperaturegradient is used in place of a denaturing gradient to identifydifferences in the mobility of control and sample DNA. See, e.g.,Rosenbaum and Reissner, 1987. Biophys. Chem. 265: 12753.

[0367] Examples of other techniques for detecting point mutationsinclude, but are not limited to, selective oligonucleotidehybridization, selective amplification, or selective primer extension.For example, oligonucleotide primers may be prepared in which the knownmutation is placed centrally and then hybridized to target DNA underconditions that permit hybridization only if a perfect match is found.See, e.g., Saiki, et al., 1986. Nature 324: 163; Saiki, et al., 1989.Proc. Natl. Acad. Sci. USA 86: 6230. Such allele specificoligonucleotides are hybridized to PCR amplified target DNA or a numberof different mutations when the oligonucleotides are attached to thehybridizing membrane and hybridized with labeled target DNA.

[0368] Alternatively, allele specific amplification technology thatdepends on selective PCR amplification may be used in conjunction withthe instant invention. Oligonucleotides used as primers for specificamplification may carry the mutation of interest in the center of themolecule (so that amplification depends on differential hybridization;see, e.g., Gibbs, et al., 1989. Nucl. Acids Res. 17: 2437-2448) or atthe extreme 3′-terminus of one primer where, under appropriateconditions, mismatch can prevent, or reduce polymerase extension (see,e.g., Prossner, 1993. Tibtech. 11: 238). In addition it may be desirableto introduce a novel restriction site in the region of the mutation tocreate cleavage-based detection. See, e.g., Gasparini, et al., 1992.Mol. Cell Probes 6: 1. It is anticipated that in certain embodimentsamplification may also be performed using Taq ligase for amplification.See, e.g., Barany, 1991. Proc. Natl. Acad. Sci. USA 88: 189. In suchcases, ligation will occur only if there is a perfect match at the3′-terminus of the 5′ sequence, making it possible to detect thepresence of a known mutation at a specific site by looking for thepresence or absence of amplification.

[0369] The methods described herein may be performed, for example, byutilizing pre-packaged diagnostic kits comprising at least one probenucleic acid or antibody reagent described herein, which may beconveniently used, e.g., in clinical settings to diagnose patientsexhibiting symptoms or family history of a disease or illness involvingan MEMX gene.

[0370] Furthermore, any cell type or tissue, preferably peripheral bloodleukocytes, in which MEMX is expressed may be utilized in the prognosticassays described herein. However, any biological sample containingnucleated cells may be used, including, for example, buccal mucosalcells.

[0371] III. Pharmacogenomics

[0372] Agents, or modulators that have a stimulatory or inhibitoryeffect on MEMX activity (e.g., MEMX gene expression), as identified by ascreening assay described herein can be administered to individuals totreat (prophylactically or therapeutically) disorders associated withaberrant MEMX activity. Such disorders for MEM1 include immunologicalconditions, viral infections, neurological disorders, Alzheimer's orParkinson's Diseases, cancer (e.g., breast or neuroblastoma),nephrology, and female reproductive health. Such disorders for MEM4include those involving the lung and/or brain (e.g., schizophrenia, orneuronal damage following head injury). Disorders for MEM5 include heartand other muscular disorders (e.g., arrhythmial), clotting deficiencies,and cobalamine deficiencies (e.g., pernicious anemia). Such disordersfor MEM6 include those originating in dysregulation of glycogenmetabolism (e.g., diabetes). Such disorders for MEM7 and MEM8 includevision-related disorders (e.g., keratomalacia), cancer, and otherneoplastic pathologies.

[0373] In conjunction with such treatment, the pharmacogenomics (i.e.,the study of the relationship between an individual's genotype and thatindividual's response to a foreign compound or drug) of the individualmay be considered. Differences in metabolism of therapeutics can lead tosevere toxicity or therapeutic failure by altering the relation betweendose and blood concentration of the pharmacologically active drug. Thus,the pharmacogenomics of the individual permits the selection ofeffective agents (e.g., drugs) for prophylactic or therapeutictreatments based on a consideration of the individual's genotype. Suchpharmacogenomics can further be used to determine appropriate dosagesand therapeutic regimens. Accordingly, the activity of MEMX protein,expression of MEMX nucleic acid, or mutation content of MEMX genes in anindividual can be determined to thereby select appropriate agent(s) fortherapeutic or prophylactic treatment of the individual.

[0374] Pharmacogenomics deals with clinically significant hereditaryvariations in the response to drugs due to altered drug disposition andabnormal action in affected persons. See e.g., Eichelbaum, 1996. Clin.Exp. Pharmacol. Physiol., 23: 983-985; Linder, 1997. Clin. Chem., 43:254-266. In general, two types of pharmacogenetic conditions can bedifferentiated. Genetic conditions transmitted as a single factoraltering the way drugs act on the body (altered drug action) or geneticconditions transmitted as single factors altering the way the body actson drugs (altered drug metabolism). These pharmacogenetic conditions canoccur either as rare defects or as polymorphisms. For example,glucose-6-phosphate dehydrogenase (G6PD) deficiency is a commoninherited enzymopathy in which the main clinical complication ishemolysis after ingestion of oxidant drugs (anti-malarials,sulfonamides, analgesics, nitrofurans) and consumption of fava beans.

[0375] As an illustrative embodiment, the activity of drug metabolizingenzymes is a major determinant of both the intensity and duration ofdrug action. The discovery of genetic polymorphisms of drug metabolizingenzymes (e.g., N-acetyltransferase 2 (NAT 2) and cytochrome P450 enzymesCYP2D6 and CYP2C19) has provided an explanation as to why some patientsdo not obtain the expected drug effects or show exaggerated drugresponse and serious toxicity after taking the standard and safe dose ofa drug. These polymorphisms are expressed in two phenotypes in thepopulation, the extensive metabolizer (EM) and poor metabolizer (PM).The prevalence of PM is different among different populations. Forexample, the gene coding for CYP2D6 is highly polymorphic and severalmutations have been identified in PM, which all lead to the absence offunctional CYP2D6. Poor metabolizers of CYP2D6 and CYP2C19 quitefrequently experience exaggerated drug response and side effects whenthey receive standard doses. If a metabolite is the active therapeuticmoiety, PM show no therapeutic response, as demonstrated for theanalgesic effect of codeine mediated by its CYP2D6-formed metabolitemorphine. At the other extreme are the so called ultra-rapidmetabolizers who do not respond to standard doses. Recently, themolecular basis of ultra-rapid metabolism has been identified to be dueto CYP2D6 gene amplification.

[0376] Thus, the activity of MEMX protein, expression of MEMX nucleicacid, or mutation content of MEMX genes in an individual can bedetermined to thereby select appropriate agent(s) for therapeutic orprophylactic treatment of the individual. In addition, pharmacogeneticstudies can be used to apply genotyping of polymorphic alleles encodingdrug-metabolizing enzymes to the identification of an individual's drugresponsiveness phenotype. This knowledge, when applied to dosing or drugselection, can avoid adverse reactions or therapeutic failure and thusenhance therapeutic or prophylactic efficiency when treating a subjectwith an MEMX modulator, such as a modulator identified by one of theexemplary screening assays described herein.

[0377] IV. Monitoring of Effects During Clinical Trials

[0378] Monitoring the influence of agents (e.g., drugs, compounds) onthe expression or activity of MEMX (e.g., the ability to modulateaberrant cell proliferation and/or differentiation) can be applied notonly in basic drug screening, but also in clinical trials. For example,the effectiveness of an agent determined by a screening assay asdescribed herein to increase MEMX gene expression, protein levels, orupregulate MEMX activity, can be monitored in clinical trails ofsubjects exhibiting decreased MEMX gene expression, protein levels, ordown-regulated MEMX activity. Alternatively, the effectiveness of anagent determined by a screening assay to decrease MEMX gene expression,protein levels, or down-regulate MEMX activity, can be monitored inclinical trails of subjects exhibiting increased MEMX gene expression,protein levels, or up-regulated MEMX activity. In such clinical trials,the expression or activity of MEMX and, preferably, other genes thathave been implicated in, for example, a cellular proliferation or immunedisorder can be used as a “read out” or markers of the immuneresponsiveness of a particular cell.

[0379] By way of example, and not of limitation, genes, including MEMX,that are modulated in cells by treatment with an agent (e.g., compound,drug or small molecule) that modulates MEMX activity (e.g., identifiedin a screening assay as described herein) can be identified. Thus, tostudy the effect of agents on cellular proliferation disorders, forexample, in a clinical trial, cells can be isolated and RNA prepared andanalyzed for the levels of expression of MEMX and other genes implicatedin the disorder. The levels of gene expression (i.e., a gene expressionpattern) can be quantified by Northern blot analysis or RT-PCR, asdescribed herein, or alternatively by measuring the amount of proteinproduced, by one of the methods as described herein, or by measuring thelevels of activity of MEMX or other genes. In this manner, the geneexpression pattern can serve as a marker, indicative of thephysiological response of the cells to the agent. Accordingly, thisresponse state may be determined before, and at various points during,treatment of the individual with the agent.

[0380] In one embodiment, the invention provides a method for monitoringthe effectiveness of treatment of a subject with an agent (e.g., anagonist, antagonist, protein, peptide, peptidomimetic, nucleic acid,small molecule, or other drug candidate identified by the screeningassays described herein) comprising the steps of (i) obtaining apre-administration sample from a subject prior to administration of theagent; (ii) detecting the level of expression of an MEMX protein, mRNA,or genomic DNA in the pre-administration sample; (iii) obtaining one ormore post-administration samples from the subject; (iv) detecting thelevel of expression or activity of the MEMX protein, mRNA, or genomicDNA in the post-administration samples; (v) comparing the level ofexpression or activity of the MEMX protein, mRNA, or genomic DNA in thepre-administration sample with the MEMX protein, mRNA, or genomic DNA inthe post administration sample or samples; and (vi) altering theadministration of the agent to the subject accordingly. For example,increased administration of the agent may be desirable to increase theexpression or activity of MEMX to higher levels than detected, i.e., toincrease the effectiveness of the agent. Alternatively, decreasedadministration of the agent may be desirable to decrease expression oractivity of MEMX to lower levels than detected, i.e., to decrease theeffectiveness of the agent.

[0381] Methods of Treatment

[0382] The invention provides for both prophylactic and therapeuticmethods of treating a subject at risk of (or susceptible to) a disorderor having a disorder associated with aberrant MEMX expression oractivity. For example, disorders associated with aberrant MEM1expression of activity include, but are not limited to, viralinfections, neurological disorders (e.g., Alzheimer's disease orParkinson's disease), cancer (e.g., breast or neuroblastoma), andvarious renal disorders. Disorders associated with aberrant MEM2, MEM3,and MEM4 expression of activity include, but are not limited to,psychiatric diseases (e.g., schizophrenia) or reducing neuronal damagefollowing head injury. Disorders associated with aberrant MEM5expression include, but are not limited to, heart and other musculardisorders (e.g., arrhythmic disorders), clotting Factor XI in clottingdeficiencies, and cobalamin-deficiencies (e.g., pernicious anemia).Disorders associated with aberrant MEM6 expression include, but are notlimited to, glycogen-metabolism-related disorders (e.g., diabetes andrelated disorders). Disorders associated with aberrant MEM7 and MEM8expression include, but are not limited to, vision-related disorders(e.g., keratomalacia) and cancer and/or similar neoplastic pathologies.

[0383] These methods of treatment will be discussed more fully, below.

[0384] I. Disease and Disorders

[0385] Diseases and disorders that are characterized by increased(relative to a subject not suffering from the disease or disorder)levels or biological activity may be treated with Therapeutics thatantagonize (i.e., reduce or inhibit) activity. Therapeutics thatantagonize activity may be administered in a therapeutic or prophylacticmanner. Therapeutics that may be utilized include, but are not limitedto: (i) an aforementioned peptide, or analogs, derivatives, fragments orhomologs thereof, (ii) antibodies to an aforementioned peptide; (iii)nucleic acids encoding an aforementioned peptide; (iv) administration ofantisense nucleic acid and nucleic acids that are “dysfunctional” (i.e.,due to a heterologous insertion within the coding sequences of codingsequences to an aforementioned peptide) that are utilized to “knockout”endogenous function of an aforementioned peptide by homologousrecombination (see, e.g., Capecchi, 1989. Science 244: 1288-1292); or(v) modulators (i.e., inhibitors, agonists and antagonists, includingadditional peptide mimetic of the invention or antibodies specific to apeptide of the invention) that alter the interaction between anaforementioned peptide and its binding partner.

[0386] Diseases and disorders that are characterized by decreased(relative to a subject not suffering from the disease or disorder)levels or biological activity may be treated with Therapeutics thatincrease (i.e., are agonists to) activity. Therapeutics that upregulateactivity may be administered in a therapeutic or prophylactic manner.Therapeutics that may be utilized include, but are not limited to, anaforementioned peptide, or analogs, derivatives, fragments or homologsthereof, or an agonist that increases bioavailability.

[0387] Increased or decreased levels can be readily detected byquantifying peptide and/or RNA, by obtaining a patient tissue sample(e.g., from biopsy tissue) and assaying it in vitro for RNA or peptidelevels, structure and/or activity of the expressed peptides (or mRNAs ofan aforementioned peptide). Methods that are well-known within the artinclude, but are not limited to, immunoassays (e.g., by Western blotanalysis, immunoprecipitation followed by sodium dodecyl sulfate (SDS)polyacrylamide gel electrophoresis, immunocytochemistry, etc.) and/orhybridization assays to detect expression of mRNAs (e.g., Northernassays, dot blots, in situ hybridization, and the like).

[0388] I. Prophylactic Methods

[0389] In one aspect, the invention provides a method for preventing, ina subject, a disease or condition associated with an aberrant MEMXexpression or activity, by administering to the subject an agent thatmodulates MEMX expression or at least one MEMX activity. Subjects atrisk for a disease that is caused or contributed to by aberrant MEMXexpression or activity can be identified by, for example, any or acombination of diagnostic or prognostic assays as described herein.Administration of a prophylactic agent can occur prior to themanifestation of symptoms characteristic of the MEMX aberrancy, suchthat a disease or disorder is prevented or, alternatively, delayed inits progression. Depending upon the type of MEMX aberrancy, for example,an MEMX agonist or MEMX antagonist agent can be used for treating thesubject. The appropriate agent can be determined based on screeningassays described herein. The prophylactic methods of the invention arefurther discussed in the following subsections.

[0390] II. Therapeutic Methods

[0391] Another aspect of the invention pertains to methods of modulatingMEMX expression or activity for therapeutic purposes. The modulatorymethod of the invention involves contacting a cell with an agent thatmodulates one or more of the activities of MEMX protein activityassociated with the cell. An agent that modulates MEMX protein activitycan be an agent as described herein, such as a nucleic acid or aprotein, a naturally-occurring cognate ligand of a MEMX protein, apeptide, a MEMX peptidomimetic, or other small molecule. In oneembodiment, the agent stimulates one or more MEMX protein activity.Examples of such stimulatory agents include active MEMX protein and anucleic acid molecule encoding MEMX that has been introduced into thecell. In another embodiment, the agent inhibits one or more MEMX proteinactivity. Examples of such inhibitory agents include antisense MEMXnucleic acid molecules and anti-MEMX antibodies. These modulatorymethods can be performed in vitro (e.g., by culturing the cell with theagent) or, alternatively, in vivo (e.g., by administering the agent to asubject). As such, the invention provides methods of treating anindividual afflicted with a disease or disorder characterized byaberrant expression or activity of a MEMX protein or nucleic acidmolecule. In one embodiment, the method involves administering an agent(e.g., an agent identified by a screening assay described herein), orcombination of agents that modulates (e.g., up-regulates ordown-regulates) MEMX expression or activity. In another embodiment, themethod involves administering a MEMX protein or nucleic acid molecule astherapy to compensate for reduced or aberrant MEMX expression oractivity.

[0392] Stimulation of MEMX activity is desirable in situations in whichMEMX is abnormally down-regulated and/or in which increased MEMXactivity is likely to have a beneficial effect. One example of such asituation is where a subject has a disorder characterized by aberrantcell proliferation and/or differentiation (e.g., cancer or immuneassociated ). Another example of such a situation is where the subjecthas an immunodeficiency disease (e.g., AIDS).

[0393] Antibodies of the invention, including polyclonal, monoclonal,humanized and fully human antibodies, may used as therapeutic agents.Such agents will generally be employed to treat or prevent a disease orpathology in a subject. An antibody preparation, preferably one havinghigh specificity and high affinity for its target antigen, isadministered to the subject and will generally have an effect due to itsbinding with the target. Such an effect may be one of two kinds,depending on the specific nature of the interaction between the givenantibody molecule and the target antigen in question. In the firstinstance, administration of the antibody may abrogate or inhibit thebinding of the target with an endogenous ligand to which it naturallybinds. In this case, the antibody binds to the target and masks abinding site of the naturally occurring ligand, wherein the ligandserves as an effector molecule. Thus the receptor mediates a signaltransduction pathway for which ligand is responsible.

[0394] Alternatively, the effect may be one in which the antibodyelicits a physiological result by virtue of binding to an effectorbinding site on the target molecule. In this case the target, a receptorhaving an endogenous ligand which may be absent or defective in thedisease or pathology, binds the antibody as a surrogate effector ligand,initiating a receptor-based signal transduction event by the receptor.

[0395] A therapeutically effective amount of an antibody of theinvention relates generally to the amount needed to achieve atherapeutic objective. As noted above, this may be a binding interactionbetween the antibody and its target antigen that, in certain cases,interferes with the functioning of the target, and in other cases,promotes a physiological response. The amount required to beadministered will furthermore depend on the binding affinity of theantibody for its specific antigen, and will also depend on the rate atwhich an administered antibody is depleted from the free volume othersubject to which it is administered. Common ranges for therapeuticallyeffective dosing of an antibody or antibody fragment of the inventionmay be, by way of non-limiting example, from about 0.1 mg/kg body weightto about 50 mg/kg body weight. Common dosing frequencies may range, forexample, from twice daily to once a week.

[0396] III. Determination of the Biological Effect of the Therapeutic

[0397] In various embodiments of the invention, suitable in vitro or invivo assays are performed to determine the effect of a specificTherapeutic and whether its administration is indicated for treatment ofthe affected tissue.

[0398] In various specific embodiments, in vitro assays may be performedwith representative cells of the type(s) involved in the patient'sdisorder, to determine if a given Therapeutic exerts the desired effectupon the cell type(s). Compounds for use in therapy may be tested insuitable animal model systems including, but not limited to rats, mice,chicken, cows, monkeys, rabbits, and the like, prior to testing in humansubjects. Similarly, for in vivo testing, any of the animal model systemknown in the art may be used prior to administration to human subjects.

SPECIFIC EXAMPLES Example 1 Real Time Quantitative (RTQ) PCR Evaluationof Expression of MEM5 in Various Cells and Tissues

[0399] The quantitative expression of MEM5 (Internal Identification16418841) was assessed in normal and tumor samples by real timequantitative PCR (TAQMAN®) performed on a Perkin-Elmer Biosystems ABIPRISMS 7700 Sequence Detection System. In the Tables contained withinthis Example, the following abbreviations are used: ca. = carcinoma,squam = squamous, * = established from metastasis, pl. eff = pl effusion= pleural effusion, met = metastasis, glio = glioma, s cell var = smallcell variant, astro = astrocytoma, non-s = non-sm = non-small, neuro =neuroblastoma

[0400] 96 RNA samples were normalized to internal standards such asβ-actin and GAPDH. RNA (˜50 ng total or ˜1 ng polyA+) was converted tocDNA using the TAQMAN® Reverse Transcription Reagents Kit (PEBiosystems; Foster City, Calif.; Catalog No. N808-0234) and randomhexamers according to the manufacturer's protocol. Reactions wereperformed in 20 μl and incubated for 30 min. at 48° C. cDNA (5 μl) wasthen transferred to a separate plate for the TAQMAN® reaction usinginternal standards such as β-actin and GAPDH TAQMAN® Assay Reagents (PEBiosystems; Catalog Nos. 4310881 E and 4310884E, respectively) andTAQMAN® Universal PCR Master Mix (PE Biosystems; Catalog No. 4304447)according to the manufacturer's protocol. Reactions were performed in 25μl total reaction volume using the following parameters: 2 minutes at50° C.; 10 minutes at 95° C.; 15 seconds at 95° C.; and I minute at 60°C. (40 cycles). Results were recorded as CT values (cycle at which agiven sample crosses a threshold level of fluorescence) using a logscale, with the difference in RNA concentration between a given sampleand the sample with the lowest CT value being represented as 2^(δCT).The percent relative expression is then obtained by taking thereciprocal of this RNA difference and multiplying by 100. The average CTvalues obtained for β-actin and GAPDH were used to normalize RNAsamples. The RNA sample generating the highest CT value required nofurther diluting, while all other samples were diluted relative to thissample according to their β-actin/GAPDH average CT values.

[0401] Normalized RNA (5 μl) was converted to cDNA and analyzed viaTAQMAN® using One Step RT-PCR Master Mix Reagents (PE Biosystems;Catalog No. 4309169) and gene-specific primers according to themanufacturer's instructions. Probes and primers were designed for eachassay according to Perkin Elmer Biosystem's Primer Express Softwarepackage (Version I for Apple Computer's Macintosh Power PC) or a similaralgorithm using the target sequence as input. Default settings were usedfor reaction conditions and the following parameters were set beforeselecting primers: primer concentration=250 nM, primer meltingtemperature (T_(m)) range=58°-60° C., primer optimal Tm=59° C., maximumprimer difference=2° C., probe does not have 5′ G, probe Tm must be 10°C. greater than primer T_(m), amplicon size 75 bp to 100 bp. The probesand primers selected (see below) were synthesized by Synthegen (Houston,Tex., USA). Probes were double purified by HPLC to remove uncoupled dyeand evaluated by mass spectroscopy to verify coupling of reporter andquencher dyes to the 5′- and 3′-termini of the probe, respectively.Their final concentrations were: forward and reverse primers=900 nMeach, and probe=200nM.

[0402] The following PCR conditions were utilized. Normalized RNA fromeach tissue and each cell line was spotted in each well of a 96 well PCRplate (Perkin Elmer Biosystems). PCR cocktails including two probes (aprobe specific for the target clone and another gene-specific probemultiplexed with the target probe) were set up using 1×TaqMan™ PCRMaster Mix for the PE Biosystems 7700, with 5 mM MgCl2, dNTPs (dA, dG,dC, dU at 1:1:1:2 ratios), 0.25 U/mi AmpliTaq Gold™ (PE Biosystems), and0.4 U/μl RNase inhibitor, and 0.25 U/μl reverse transcriptase. Reversetranscription was performed at 48° C. for 30 minutes followed byamplification/PCR cycles as follows: 95° C. 10 minutes; then 40 cyclesof 95° C. for 15 seconds; 60° C. for 1 minute.

[0403] Two sample panels were employed in the present Example. Panel 1is a 96 well plate (usually 2 control wells and 94 test samples) whosewells are contain RNA or cDNA isolated from various human cell linesthat have been established from human malignant tissues (i.e., tumors).These cell lines have been extensively characterized by investigators inboth academia and the commercial sector regarding their tumorgenicity,metastatic potential, drug resistance, invasive potential and othercancer-related properties. They serve as suitable tools for pre-clinicalevaluation of anti-cancer agents and promising therapeutic strategies.RNA from these various human cancer cell lines was isolated by andprocured from the Developmental Therapeutic Branch (DTB) of the NationalCancer Institute (USA). Basic information regarding their biologicalbehavior, gene expression, and resistance to various cytotoxic agentsare provided by the DTB (http://dtp.nci.nih.gov/).

[0404] In addition, RNA or cDNA was obtained from various human tissuesderived from human autopsies performed on deceased elderly people orsudden death victims (accidents, etc.). These tissues were ascertainedto be free of disease and were purchased from various high qualitycommercial sources such as Clontech, Research Genetics, and Invitrogen.

[0405] RNA integrity from all samples was controlled for quality byvisual assessment of agarose gel electrophoresis using 28 S and 18 Sribosomal RNA (rRNA) staining intensity ratio as a guide (2:1 to 2.5:128 S:18 S rRNA ratio) and the assuring the absence of low molecularweight RNAs indicative of degradation products.

[0406] Panel 2 is a 96 well plate (usually 2 control wells and 94 testsamples) containing RNA or cDNA isolated from human tissue procured bysurgeons working in close cooperation with the National CancerInstitute's Cooperative Human Tissue Network (CHTN) or the NationalDisease Research Initiative (NDRI). The tissues procured are derivedfrom human malignancies and in cases where indicated many malignanttissues have “matched margins”. The tumor tissue and the “matchedmargins” are evaluated by two independent pathologists (the surgicalpathologists and again by a pathologists at NDRI or CHTN). This analysisprovides a gross histopathological assessment of tumor differentiationgrade. Moreover, most samples include the original surgical pathologyreport that provides information regarding the clinical stage of thepatient. These matched margins are taken from the tissue surrounding(i.e., immediately proximal) to the zone of surgery (designated “NAT”,for normal adjacent tissue). In addition, RNA or cDNA was obtained fromvarious human tissues derived from human autopsies performed on deceasedelderly people or sudden death victims (accidents, etc.). These tissuewere ascertained to be free of disease and were purchased from varioushigh quality commercial sources such as Clontech, Research Genetics, andInvitrogen.

[0407] Again, RNA integrity from all samples was controlled for qualityby visual assessment of agarose gel electrophoresis using 28 S and 18 SrRNA staining intensity ratio as a guide (2:1 to 2.5:1 28 S:18 S ratio)and by assuring the absence of low molecular weight RNAs indicative ofdegradation products. Samples are quality controlled for genomic DNAcontamination by reactions run in the absence of reverse transcriptaseusing probe and primer sets designed to amplify across the span of asingle exon.

[0408] The following RTQ PCR of the MEM5 sequence (Internal Designation16418841) utilzing Panel 1, is shown in Table 3, using the primer-probeset Ag765 designated in Table 2. TABLE 2 SEQ Start ID Primers SequencesLength Position NO: Forward 5′-CCAACGTGAAGGGAGCTATAT-3′ 21 923 17 ProbeTET-5′-TGCTGACACCACTACACATGTCACAA-3′-TAMRA 26 953 18 Reverse5′-CCAGCCCCTAAAATTCTCATC-3′ 21 986 19

[0409] TABLE 3 Rel. Expr., Rel. Expr., Rel. Expr., Rel. Expr., Rel.Expr., Rel. Expr., % % % % % % Cell source 1.2tm717t 1.2tm917t 1.2tm971tCell source 1.2tm717t 1.2tm917t 1.2tm971t Endothelial cells 22.5 22.522.1 Renal ca. 786-0 17.6 17.6 5.3 Endothelial cells 2.0 2.0 1.3 Renalca. A498 27.2 27.2 22.1 (treated) Pancreas 27.7 27.7 32.5 Renal ca. RXF393 2.8 2.8 2.5 Pancreatic ca. 11.8 11.8 3.9 Renal ca. ACHN 8.7 8.7 6.9CAPAN 2 Adrenal Gland (new 12.5 12.5 14.9 Renal ca. UO-31 16.6 16.6 4.5lot*) Thyroid 24.8 24.8 18.7 Renal ca. TK-10 20.3 20.3 9.1 Salivarygland 35.1 35.1 33.9 Liver 6.8 6.8 6.9 Pituitary gland 33.7 33.7 44.1Liver (fetal) 12.9 12.9 12.8 Brain (fetal) 1.7 1.7 3.2 Liver ca. 8.8 8.83.2 (hepatoblast) HepG2 Brain (whole) 11.9 11.9 13.0 Lung 11.8 11.8 10.7Brain (amygdala) 2.9 2.9 4.4 Lung (fetal) 10.9 10.9 12.6 Brain(cerebellum) 6.6 6.6 10.0 Lung ca. (small cell) 26.2 26.2 16.5 LX-1Brain (hippocampus) 6.7 6.7 8.9 Lung ca. (small cell) 28.7 28.7 9.2NCI-H69 Brain (thalamus) 6.1 6.1 5.5 Lung ca. (s. cell var.) 12.5 12.57.7 SHP-77 Cerebral Cortex 7.6 7.6 0.0 Lung ca. (large 23.7 23.7 17.1cell) NCI-H460 Spinal cord 7.1 7.1 10.7 Lung ca. (non-sm. 12.6 12.6 6.5cell) A549 CNS ca. (glio/astro) 22.9 22.9 22.9 Lung ca. (non-s. cell)7.3 7.3 6.0 U87-MG NCI-H23 CNS ca. (glio/astro) 33.5 33.5 25.9 Lung ca.(non-s. cell) 11.5 11.5 0.0 U-118-MG HOP-62 CNS ca. (astro) 7.8 7.8 6.0Lung ca. (non-s. cl) 44.1 44.1 24.7 SW1783 NCI-H522 CNS ca.* (neuro;26.1 26.1 23.3 Lung ca. (squam.) 15.9 15.9 8.1 met) SK-N-AS SW 900 CNSca. (astro) SF- 16.7 16.7 11.6 Lung ca. (squam.) 21.6 21.6 16.4 539NCI-H596 CNS ca. (astro) 19.6 19.6 12.5 Mammary gland 24.8 24.8 24.7SNB-75 CNS ca. (glio) 65.1 65.1 27.0 Breast ca.* (pl. 11.7 11.7 7.6SNB-19 effusion) MCF-7 CNS ca. (glio) U251 17.2 17.2 9.0 Breast ca.*(pl. ef) 14.3 14.3 13.4 MDA-MB-231 CNS ca. (glio) SF- 22.1 22.1 20.0Breast ca.* (pl. 11.0 11.0 12.3 295 effusion) T47D Heart 57.8 57.8 57.4Breast ca. BT-549 21.2 21.2 15.3 Skeletal Muscle 100.0 100.0 100.0Breast ca. MDA-N 10.8 10.8 13.4 (new lot*) Bone marrow 7.2 7.2 7.2 Ovary2.2 2.2 0.3 Thymus 5.6 5.6 4.2 Ovarian ca. 15.5 15.5 12.5 OVCAR-3 Spleen6.0 6.0 7.4 Ovarian ca. 5.0 5.0 5.0 OVCAR-4 Lymph node 7.0 7.0 7.7Ovarian ca. 12.2 12.2 6.4 OVCAR-5 Colorectal 2.2 2 2 1.3 Ovarian ca.15.3 15.3 5.6 OVCAR-8 Stomach 12.4 12.4 15.8 Ovarian ca. IGROV-1 20.520.5 12.9 Small intestine 23.5 23.5 17.8 Ovarian ca.* 30.6 30.6 26.6(ascites) SK-OV-3 Colon ca. SW480 15.6 15.6 6.5 Uterus 92 9.2 6.3 Colonca.* (SW480 38.4 38.4 15.9 Placenta 13.7 13.7 15.2 met) SW620 Colon ca.HT29 8.5 8.5 3.1 Prostate 20.6 20.6 16.6 Colon ca. HCT-116 20.9 20.910.5 Prostate ca.* (bone 23.7 23.7 15.5 met) PC-3 Colon ca. CaCo-2 11.511 5 4.5 Testis 29.5 29.5 23.8 83219 CC Well to 3.9 3.9 0.4 Melanoma12.0 12.0 8.4 Mod Diff Hs688(A).T (ODO3866) Colon ca. HCC-2998 35.4 35.421.6 Melanoma* (met) 15.9 15.9 11.0 Hs688(B).T Gastric ca.* (liver 25.525.5 21.0 Melanoma UACC-62 1.9 1.9 1.0 met) NCI-N87 Bladder 23.0 23.022.5 Melanoma M14 14.4 14.4 1.7 Trachea 9.4 9.4 10.7 Melanoma LOX 8.38.3 2.1 IMVI Kidney 12.9 12.9 13.1 Melanoma* (met) 14.1 14.1 11.2SK-MEL-5 Kidney (fetal) 19.0 19.0 8.3 Adipose 2.1 2.1 1.0

[0410] The results in Table 3 show that the sequence of MEM5 isexpressed in a wide variety of normal and cancer cell lines. Withrelation to normal tissues, it is more highly expressed in certain braintumors such as CNS ca. (glio) SNB-19, colon cancer such as Colon ca.*(SW480 met)SW620, and lung cancer such as Lung ca. (non-s.cl) NCI-H522.

[0411] Additional results for MEM5 which were obtained using Panel 2 areshown in Table 4. TABLE 4 Rel. Expr., % Tissue source 2tm972t 83786Kidney Ca, Nuclear grade 2 (OD04338) 15.4 83219 CC Well to Mod Diff(ODO3866) 2.1 83220 CC NAT (ODO3866) 11.7 83221 CC Gr.2 rectosigmoid(ODO3868) 11.7 83222 CC NAT (ODO3868) 3.4 83235 CC Mod Diff (ODO3920)28.9 83236 CC NAT (ODO3920) 30.8 83237 CC Gr.2 ascend colon (ODO3921)10.4 83238 CC NAT (ODO3921) 1.6 83239 Lung Met to Muscle (ODO4286) 1.583240 Muscle NAT (ODO4286) 18.4 83241 CC from Partial Hepatectomy(ODO4309) 9.6 83242 Liver NAT (ODO4309) 22.1 83255 Ocular Mel Met toLiver (ODO4310) 54.7 83256 Liver NAT (ODO4310) 25.2 83787 Kidney NAT(ODO4338) 32.8 83788 Kidney Ca Nuclear grade 1/2 (OD04339) 59.1 83789Kidney NAT (OD04339) 50.4 83790 Kidney Ca, Clear cell type (OD04340)100.0 83791 Kidney NAT (OD04340) 39.2 83792 Kidney Ca, Nuclear grade 3(OD04348) 24.2 83793 Kidney NAT (OD04348) 25.4 84136 Lung MalignantCancer (OD03126) 5.5 84137 Lung NAT (OD03126) 9.7 84138 Lung NAT(OD04321) 4.8 84139 Melanoma Mets to Lung (OD04321) 26.2 84140 ProstateCancer (OD04410) 26.8 84141 Prostate NAT (OD04410) 41.8 84871 LungCancer (OD04404) 2.0 84872 Lung NAT (OD04404) 0.0 84875 Lung Cancer(OD04565) 55.9 84877 Breast Cancer (OD04566) 7.2 85950 Lung Cancer(OD04237-01) 7.0 85970 Lung NAT (OD04237-02) 12.2 85973 Kidney Cancer(OD04450-01) 20.6 85974 Kidney NAT (OD04450-03) 40.6 85975 Breast Cancer(OD04590-01) 31.6 85976 Breast Cancer Mets (OD04590-03) 39.5 87070Breast Cancer Metastasis (OD04655-05) 26.6 87071 Bladder Cancer(OD04718-01) 28.3 87072 Bladder Normal Adjacent (OD04718-03) 1.8 87073Prostate Cancer (OD04720-01) 52.5 87074 Prostate NAT (OD04720-02) 6.287472 Colon mets to lung (OD04451-01) 6.7 87473 Lung NAT (OD04451-02)4.3 87474 Kidney Cancer (OD04622-01) 7.5 87475 Kidney NAT (OD04622-03)1.7 87492 Ovary Cancer (OD04768-07) 78.5 87493 Ovary NAT (OD04768-08)11.7 Bladder Cancer INVITROGEN A302173 7.7 Bladder Cancer ResearchGenetics RNA 1023 0.0 Breast Cancer Clontech 9100266 2.8 Breast CancerINVITROGEN A209073 1.9 Breast Cancer Res. Gen. 1024 21.8 Breast NATClontech 9100265 0.1 Breast NAT INVITROGEN A2090734 5.1 GENPAK BreastCancer 064006 17.3 Gastric Cancer Clontech 9060395 14.8 Gastric CancerClontech 9060397 3.0 Gastric Cancer GENPAK 064005 48.6 Kidney CancerClontech 8120607 0.0 Kidney Cancer Clontech 8120613 0.3 Kidney CancerClontech 9010320 0.4 Kidney NAT Clontech 8120608 0.1 Kidney NAT Clontech8120614 0.0 Kidney NAT Clontech 9010321 0.1 Liver Cancer GENPAK 06400323.8 Liver Cancer Research Genetics RNA 1025 0 8 Liver Cancer ResearchGenetics RNA 1026 0.1 NAT Stomach Clontech 9060359 14.3 NAT StomachClontech 9060394 11.4 NAT Stomach Clontech 9060396 5.4 Normal BladderGENPAK 061001 18.8 Normal Breast GENPAK 061019 5.4 Normal Colon GENPAK061003 17.2 Normal Kidney GENPAK 061008 11.8 Normal Liver GENPAK 06100926.4 Normal Lung GENPAK 061010 10.9 Normal Ovary Res. Gen. 0.6 NormalProstate Clontech A+ 6546-1 3.9 Normal Stomach GENPAK 061017 12.9 NormalThyroid Clontech A+ 6570-1** 10.3 Normal Uterus GENPAK 061018 12.6Ovarian Cancer GENPAK 064008 9.3 Paired Liver Cancer Tissue RNA 6004-T0.4 Paired Liver Cancer Tissue RNA 6005-T 1.3 Paired Liver Tissue RNA6004-N 9.0 Paired Liver Tissue Research Genetics RNA 0.8 6005-N ThyroidCancer GENPAK 064010 17.8 Thyroid Cancer INVITROGEN A302152 27.7 ThyroidNAT INVITROGEN A302153 23.2 Uterus Cancer GENPAK 064011 29.1 genomic DNAcontrol 0.3 87492 Ovary Cancer (OD04768-07) 78.5

[0412] The results shown in Table 4 indicate that MEM5 is expressedpreferentially in certain tumor samples compared to the adjacentnoncancerous tissue. These tumors include a liver metastasis, a kidneytumor, a prostate cancer, and an ovarian cancer. In addition there ishigh expression in additional tumor tissues that have no matching normaltissue in the panel.

[0413] Accordingly, the results in Tables 3 and 4 suggests that MEM5 mayserve as a diagnostic probe for certain specific cancer types.

Example 2 Real Time Quantitative (RTQ) PCR Evaluation of Expression ofMEM7 in Various Cells and Tissues

[0414] The quantitative expression of MEM7 (Internal IdentificationAC018653_A) was assessed in normal and tumor samples by real timequantitative PCR (TAQMAN®) performed on a Perkin-Elmer Biosystems ABIPRISM® 7700 Sequence Detection System. In the Tables contained withinthis Example, the following abbreviations are used: ca. = carcinoma,squam = squamous, * = established from metastasis, pl. eff = pl effusion= pleural effusion, met = metastasis, glio = glioma, s cell var = smallcell variant, astro = astrocytoma, non-s = non-sm = non-small, neuro =neuroblastoma

[0415] 96 RNA samples were normalized to internal standards such asβ-actin and GAPDH. RNA (˜50 ng total or ˜1 ng poly A+) was converted tocDNA using the TAQMAN® Reverse Transcription Reagents Kit (PEBiosystems; Foster City, Calif.; Catalog No. N808-0234) and randomhexamers according to the manufacturer's protocol. Reactions wereperformed in 20 μl and incubated for 30 min. at 48° C. cDNA (5 μl) wasthen transferred to a separate plate for the TAQMAN® reaction usinginternal standards such as β-actin and GAPDH TAQMAN® Assay Reagents (PEBiosystems; Catalog Nos. 4310881 E and 4310884E, respectively) andTAQMAN® Universal PCR Master Mix (PE Biosystems; Catalog No. 4304447)according to the manufacturer's protocol. Reactions were performed in 25μl total reaction volume using the following parameters: 2 minutes at50° C.; 10 minutes at 95° C.; 15 seconds at 95° C.; and 1 minute at 60°C. (40 cycles). Results were recorded as CT values (cycle at which agiven sample crosses a threshold level of fluorescence) using a logscale, with the difference in RNA concentration between a given sampleand the sample with the lowest CT value being represented as 2^(δCT).The percent relative expression is then obtained by taking thereciprocal of this RNA difference and multiplying by 100. The average CTvalues obtained for β-actin and GAPDH were used to normalize RNAsamples. The RNA sample generating the highest CT value required nofurther diluting, while all other samples were diluted relative to thissample according to their β-actin/GAPDH average CT values.

[0416] Normalized RNA (5 μl) was converted to cDNA and analyzed viaTAQMAN® using One Step RT-PCR Master Mix Reagents (PE Biosystems;Catalog No. 4309169) and gene-specific primers according to themanufacturer's instructions. Probes and primers were designed for eachassay according to Perkin Elmer Biosystem's Primer Express Softwarepackage (Version I for Apple Computer's Macintosh Power PC) or a similaralgorithm using the target sequence as input. Default settings were usedfor reaction conditions and the following parameters were set beforeselecting primers: primer concentration=250 nM, primer meltingtemperature (T_(m)) range=58°-60° C., primer optimal Tm=59° C., maximumprimer difference=2° C., probe does not have 5′ G, probe T_(m) must be10° C. greater than primer T_(m), amplicon size 75 bp to 100 bp. Theprobes and primers selected (see below) were synthesized by Synthegen(Houston, Tex., USA). Probes were double purified by HPLC to removeuncoupled dye and evaluated by mass spectroscopy to verify coupling ofreporter and quencher dyes to the 5′- and 3′-termini of the probe,respectively. Their final concentrations were: forward and reverseprimers=900 nM each, and probe=200 nM.

[0417] The following PCR conditions were utilized. Normalized RNA fromeach tissue and each cell line was spotted in each well of a 96 well PCRplate (Perkin Elmer Biosystems). PCR cocktails including two probes (aprobe specific for the target clone and another gene-specific probemultiplexed with the target probe) were set up using 1×TaqMan® PCRMaster Mix for the PE Biosystems 7700, with 5 mM MgCl2, dNTPs (dA, dG,dC, dU at 1:1:1:2 ratios), 0.25 U/ml AmpliTaq Gold™ (PE Biosystems), and0.4 U/μl RNase inhibitor, and 0.25 U/μl reverse transcriptase. Reversetranscription was performed at 48° C. for 30 minutes followed byamplification/PCR cycles as follows: 95° C. 10 minutes; then 40 cyclesof 95° C. for 15 seconds; 60° C. for 1 minute.

[0418] Two sample panels were employed in the present Example. Panel 1is a 96 well plate (usually 2 control wells and 94 test samples) whosewells are contain RNA or cDNA isolated from various human cell linesthat have been established from human malignant tissues (i.e., tumors).These cell lines have been extensively characterized by investigators inboth academia and the commercial sector regarding their tumorgenicity,metastatic potential, drug resistance, invasive potential and othercancer-related properties. They serve as suitable tools for pre-clinicalevaluation of anti-cancer agents and promising therapeutic strategies.RNA from these various human cancer cell lines was isolated by andprocured from the Developmental Therapeutic Branch (DTB) of the NationalCancer Institute (USA). Basic information regarding their biologicalbehavior, gene expression, and resistance to various cytotoxic agentsare provided by the DTB (http://dtp.nci.nih.gov/).

[0419] In addition, RNA or cDNA was obtained from various human tissuesderived from human autopsies performed on deceased elderly people orsudden death victims (accidents, etc.). These tissues were ascertainedto be free of disease and were purchased from various high qualitycommercial sources such as Clontech, Research Genetics, and Invitrogen.

[0420] RNA integrity from all samples was controlled for quality byvisual assessment of agarose gel electrophoresis using 28 S and 18 Sribosomal RNA (rRNA) staining intensity ratio as a guide (2:1 to 2.5:128 S:18 S rRNA ratio) and the assuring the absence of low molecularweight RNAs indicative of degradation products.

[0421] Panel 2 is a 96 well plate (usually 2 control wells and 94 testsamples) containing RNA or cDNA isolated from human tissue procured bysurgeons working in close cooperation with the National CancerInstitute's Cooperative Human Tissue Network (CHTN) or the NationalDisease Research Initiative (NDRI). The tissues procured are derivedfrom human malignancies and in cases where indicated many malignanttissues have “matched margins”. The tumor tissue and the “matchedmargins” are evaluated by two independent pathologists (the surgicalpathologists and again by a pathologists at NDRI or CHTN). This analysisprovides a gross histopathological assessment of tumor differentiationgrade. Moreover, most samples include the original surgical pathologyreport that provides information regarding the clinical stage of thepatient. These matched margins are taken from the tissue surrounding(i.e., immediately proximal) to the zone of surgery (designated “NAT”,for normal adjacent tissue). In addition, RNA or cDNA was obtained fromvarious human tissues derived from human autopsies performed on deceasedelderly people or sudden death victims (accidents, etc.). These tissuewere ascertained to be free of disease and were purchased from varioushigh quality commercial sources such as Clontech, Research Genetics, andInvitrogen.

[0422] Again, RNA integrity from all samples was controlled for qualityby visual assessment of agarose gel electrophoresis using 28 S and 18 SrRNA staining intensity ratio as a guide (2:1 to 2.5:1 28 S:18 S ratio)and by assuring the absence of low molecular weight RNAs indicative ofdegradation products. Samples are quality controlled for genomic DNAcontamination by reactions run in the absence of reverse transcriptaseusing probe and primer sets designed to amplify across the span of asingle exon.

[0423] The following RTQ PCR of the MEM7 sequence (InternalIdentification ACO18653_A) utilzing Panel 1, is shown in Table 6, usingthe primer-probe set Ag 1387 designated in Table 5. TABLE 5 StartPrimers Sequences Length Position SEQ ID NO: Forward5′-CTGAAACCTTCATCCACACAAT-3′ 22 18 20 ProbeTET-5′-TCACTGGCTACTACCGCTTTGTCTCG-3′- 26 51 21 TAMRA Reverse5′-GCAGGTAGTCCTCCATGTTCTT-3′ 22 80 22

[0424] TABLE 6 Rel. Expr., %, Cell source 1.2tm1615t Endothelial cells0.1 Endothelial cells (treated) 0.6 Pancreas 0.0 Pancreatic Ca. CAPAN 20.1 Adrenal Gland (new lot*) 0.4 Thyroid 0.0 Salavary gland 0.7Pituitary gland 0.0 Brain (fetal) 0.0 Brain (whole) 0.0 Brain (amygdala)0.0 Brain (cerebellum) 0.0 Brain (hippocampus) 0.1 Brain (thalamus) 0.1Cerebral Cortex 0.2 Spinal cord 0.0 CNS ca. (glio/astro) U87-MG 0.1 CNSca. (glio/astro) U-118-MG 0.1 CNS ca. (astro) SW1783 0.1 CNS ca.*(neuro; met) SK-N-AS 0.1 CNS ca. (astro) SF-539 0.3 CNS ca. (astro)SNB-75 0.0 CNS ca. (glio) SNB-19 0.1 CNS ca. (glio) U251 0.0 CNS ca.(glio) SF-295 0.3 Heart 0.3 Skeletal Muscle (new lot*) 0.0 Bone marrow0.2 Thymus 0.4 Spleen 1.5 Lymph node 1.3 Colorectal 0 0 Stomach 0.9Small intestine 1.8 Colon ca. SW480 0.1 Colon ca.* (SW480 met)SW620 0.3Colon ca. HT29 0.3 Colon ca. HCT-116 0.3 Colon ca. CaCo-2 0.5 83219 CCWell to Mod Diff (ODO3866) 0.2 Colon ca. HCC-2998 1.5 Gastric ca.*(liver met) NCI-N87 0.7 Bladder 2.5 Trachea 0.1 Kidney 100.0 Kidney(fetal) 0.3 Renal ca. 786-0 0.2 Renal ca. A498 0.7 Renal ca. RXF 393 0.3Renal ca. ACHN 0.7 Renal ca. UO-31 0.5 Renal ca. TK-10 0.3 Liver 12.7Liver (fetal) 1.4 Liver ca. (hepatoblast) HepG2 1.7 Lung 0.1 Lung(fetal) 0.3 Lung ca. (small cell) LX-1 1.1 Lung ca. (small cell) NCI-H690.7 Lung ca. (s.cell var.) SHP-77 0.0 Lung ca. (large cell)NCI-H460 1.7Lung ca. (non-sm. cell) A549 0.6 Lung ca. (non-s.cell) NCI-H23 1.3 Lungca (non-s. cell) HOP-62 0 6 Lung ca. (non-s.cl) NCI-H522 0.9 Lung ca.(squam.) SW 900 0.5 Lung ca. (squam.) NCI-H596 0.3 Mammary gland 0.3Breast ca.* (pl. effusion) MCF-7 0.0 Breast ca.* (pl.ef) MDA-MB-231 0.0Breast ca.* (pl. effusion) T47D 0.1 Breast ca. BT-549 0.0 Breast ca.MDA-N 0.1 Ovary 0.7 Ovarian ca. OVCAR-3 0 2 Ovarian ca. OVCAR-4 0.2Ovarian ca. OVCAR-5 1.3 Ovarian ca. OVCAR-8 0.3 Ovarian ca. IGROV-1 0.3Ovarian ca.* (ascites) SK-OV-3 0.4 Uterus 1.0 Plancenta 0.0 Prostate 0.9Prostate ca.* (bone met)PC-3 0.4 Testis 0.0 Melanoma Hs688(A).T 0.1Melanoma* (met) Hs688(B).T 0.1 Melanoma UACC-62 0 1 Melanoma M14 0 0Melanoma LOX IMVI 0.0 Melanoma* (met) SK-MEL-S 0.0 Adipose 1.7

[0425] Additionally, the expression of sequence MEM7 was also evaluatedusing the same primer-probe set, Ag1387, on Panel 2. The results areshown in Table 7. TABLE 7 Rel. Expr., %, Tissue Source 2tm515f 83786Kidney Ca, Nuclear grade 2 (OD04338) 9.9 83219 CC Well to Mod Diff(ODO3866) 3.7 83220 CC NAT (ODO3866) 3.7 83221 CC Gr.2 rectosigmoid(ODO3868) 3.4 83222 CC NAT (ODO3868) 1.3 83235 CC Mod Diff (ODO3920)10.6 83236 CC NAT (ODO3920) 5.0 83237 CC Gr.2 ascend colon (ODO3921) 4.583238 CC NAT (ODO3921) 2.3 83239 Lung Met to Muscle (ODO4286) 2.6 83240Muscle NAT (ODO4286) 6.4 83241 CC from Partial Hepatectomy (ODO4309) 7.183242 Liver NAT (ODO4309) 80.1 83255 Ocular Mel Met to Liver (ODO4310)1.7 83256 Liver NAT (ODO4310) 51.4 83787 Kidney NAT (OD04338) 11.5 83788Kidney Ca Nuclear grade 1/2 (OD04339) 26.4 83789 Kidney NAT (OD04339)10.7 83790 Kidney Ca, Clear cell type (OD04340) 12.0 83791 Kidney NAT(OD04340) 9.2 83792 Kidney Ca, Nuclear grade 3 (OD04348) 6.7 83793Kidney NAT (OD04348) 12.9 84136 Lung Malignant Cancer (OD03126) 4.784137 Lung NAT (OD03126) 9.7 84138 Lung NAT (OD04321) 4.7 84139 MelanomaMets to Lung (OD04321) 5.1 84140 Prostate Cancer (OD04410) 7.0 84141Prostate NAT (OD04410) 11.0 84871 Lung Cancer (OD04404) 4.4 84872 LungNAT (OD04404) 4.3 84875 Lung Cancer (OD04565) 9.3 84877 Breast Cancer(OD04566) 7.8 85950 Lung Cancer (OD04237-01) 6.2 85970 Lung NAT(OD04237-02) 5.1 85973 Kidney Cancer (OD04450-01) 13.4 85974 Kidney NAT(OD04450-03) 6.3 85975 Breast Cancer (OD04590-01) 6.3 85976 BreastCancer Mets (OD04590-03) 7.1 87070 Breast Cancer Metastasis (OD04655-05)9.4 87071 Bladder Cancer (OD04718-01) 5.3 87072 Bladder Normal Adjacent(OD04718-03) 4.8 87073 Prostate Cancer (OD04720-01) 13.1 87074 ProstateNAT (OD04720-02) 10.4 87472 Colon mets to lung (OD04451-01) 9.5 87473Lung NAT (OD04451 -02) 5.6 87474 Kidney Cancer (OD04622-01) 27.4 87475Kidney NAT (OD04622-03) 7.1 87492 Ovary Cancer (OD04768-07) 17.7 87493Ovary NAT (OD04768-08) 9.0 Bladder Cancer INVITROGEN A302173 7.9 BladderCancer Research Genetics RNA 1023 4.0 Breast Cancer Clontech 9100266 6.1Breast Cancer INVITROGEN A209073 6.6 Breast Cancer Res. Gen. 1024 9.9Breast NAT Clontech 9100265 3.8 Breast NAT INVITROGEN A2090734 8.0GENPAK Breast Cancer 064006 12.8 Gastric Cancer Clontech 9060395 5.8Gastric Cancer Clontech 9060397 6.3 Gastric Cancer GENPAK 064005 12.5Kidney Cancer Clontech 8120607 8.3 Kidney Cancer Clontech 8120613 1.0Kidney Cancer Clontech 9010320 3.7 Kidney NAT Clontech 8120608 4.4Kidney NAT Clontech 8120614 2.9 Kidney NAT Clontech 9010321 3.1 LiverCancer GENPAK 064003 23.5 Liver Cancer Research Genetics RNA 1025 92.7Liver Cancer Research Genetics RNA 1026 16.6 NAT Stomach Clontech9060359 7.1 NAT Stomach Clontech 9060394 7.2 NAT Stomach Clontech9060396 3.4 Normal Bladder GENPAK 061001 13.0 Normal Breast GENPAK061019 8.1 Normal Colon GENPAK 061003 5.2 Normal Kidney GENPAK 0610084.8 Normal Liver GENPAK 061009 100.0 Normal Lung GENPAK 061010 5.7Normal Ovary Res. Gen. 7.0 Normal Prostate Clontech A+ 6546-1 4.8 NormalStomach GENPAK 061017 6.4 Normal Thyroid Clontech A+ 6570-1** 6.1 NormalUterus GENPAK 061018 3.0 Ovarian Cancer GENPAK 064008 11.7 Paired LiverCancer Tissue RNA 6004-T 54.0 Paired Liver Cancer Tissue RNA 6005-T 18.4Paired Liver Tissue RNA 6004-N 26.1 Paired Liver Tissue Genetics RNA6005-N 55.1 Thyroid Cancer GENPAK 064010 3.4 Thyroid Cancer INVITROGENA302152 10.7 Thyroid NAT INVITROGEN A302153 7.7 Uterus Cancer GENPAK064011 10.6 Genomic DNA control 0.6

[0426] The results for MEM7 indicate expression primarily in normalkidney and lung tissue, and, for certain tumors but not all, in normaltissue adjacent to certain tumors in these organs. These results suggestthat MEM7 may be used to distinguish normal from cancerous tissue.

[0427] Other Embodiments

[0428] While the invention has been described in conjunction with thedetailed description thereof, the foregoing description is intended toillustrate and not limit the scope of the invention, which is defined bythe scope of the appended claims. Other aspects, advantages, andmodifications are within the scope of the following claims.

1 49 1 9045 DNA Homo sapiens CDS (1)..(9042) 1 atg gcg ccg ccg ccg ccgccc gtg ctg ccc gtg ctg ctg ctc ctg gcc 48 Met Ala Pro Pro Pro Pro ProVal Leu Pro Val Leu Leu Leu Leu Ala 1 5 10 15 gcc gcc gcc gcc ctg ccggcg atg ggg ctg cga gcg gcc gcc tgg gag 96 Ala Ala Ala Ala Leu Pro AlaMet Gly Leu Arg Ala Ala Ala Trp Glu 20 25 30 ccg cgc gta ccc ggc ggg acccgc gcc ttc gcc ctc cgg ccc ggc tgt 144 Pro Arg Val Pro Gly Gly Thr ArgAla Phe Ala Leu Arg Pro Gly Cys 35 40 45 acc tac gcg gtg ggc gcc gct tgcacg ccc cgg gcg ccg cgg gag ctg 192 Thr Tyr Ala Val Gly Ala Ala Cys ThrPro Arg Ala Pro Arg Glu Leu 50 55 60 ctg gac gtg ggc cgc gat ggg cgg ctggca gga cgt cgg cgc gtc tcg 240 Leu Asp Val Gly Arg Asp Gly Arg Leu AlaGly Arg Arg Arg Val Ser 65 70 75 80 ggc gcg ggg cgc ccg ctg ccg ctg caagtc cgc ttg gtg gcc cgc agt 288 Gly Ala Gly Arg Pro Leu Pro Leu Gln ValArg Leu Val Ala Arg Ser 85 90 95 gcc ccg acg gcg ctg agc cgc cgc ctg cgggcg cgc acg cac ctt ccc 336 Ala Pro Thr Ala Leu Ser Arg Arg Leu Arg AlaArg Thr His Leu Pro 100 105 110 ggc tgc gga gcc cgt gcc cgg ctc tgc ggaacc ggt gcc cgg ctc tgc 384 Gly Cys Gly Ala Arg Ala Arg Leu Cys Gly ThrGly Ala Arg Leu Cys 115 120 125 ggg gcg ctc tgc ttc ccc gtc ccc ggc ggctgc gcg gcc gcg cag cat 432 Gly Ala Leu Cys Phe Pro Val Pro Gly Gly CysAla Ala Ala Gln His 130 135 140 tcg gcg ctc gca gct ccg acc acc tta cccgcc tgc cgc tgc ccg ccg 480 Ser Ala Leu Ala Ala Pro Thr Thr Leu Pro AlaCys Arg Cys Pro Pro 145 150 155 160 cgc ccc agg ccc cgc tgt ccc ggc cgtccc atc tgc ctg ccg ccg ggc 528 Arg Pro Arg Pro Arg Cys Pro Gly Arg ProIle Cys Leu Pro Pro Gly 165 170 175 ggc tcg gtc cgc ctg cgt ctg ctg tgcgcc ctg cgg cgc gcg gct ggc 576 Gly Ser Val Arg Leu Arg Leu Leu Cys AlaLeu Arg Arg Ala Ala Gly 180 185 190 gcc gtc cgg gtg gga ctg gcg ctg gaggcc gcc acc gcg ggg acg ccc 624 Ala Val Arg Val Gly Leu Ala Leu Glu AlaAla Thr Ala Gly Thr Pro 195 200 205 tcc gcg tcg cca tcc cca tcg ccg cccctg ccg ccg aac ttg ccc gaa 672 Ser Ala Ser Pro Ser Pro Ser Pro Pro LeuPro Pro Asn Leu Pro Glu 210 215 220 gcc cgg gcg ggg ccg gcg cga cgg gcccgg cgg ggc acg agc ggc aga 720 Ala Arg Ala Gly Pro Ala Arg Arg Ala ArgArg Gly Thr Ser Gly Arg 225 230 235 240 ggg agc ctg aag ttt ccg atg cccaac tac cag gtg gcg ttg ttt gag 768 Gly Ser Leu Lys Phe Pro Met Pro AsnTyr Gln Val Ala Leu Phe Glu 245 250 255 aac gaa ccg gcg ggc acc ctc atcctc cag ctg cac gcg cac tac acc 816 Asn Glu Pro Ala Gly Thr Leu Ile LeuGln Leu His Ala His Tyr Thr 260 265 270 atc gag ggc gag gag gag cgc gtgagc tat tac atg gag ggg ctg ttc 864 Ile Glu Gly Glu Glu Glu Arg Val SerTyr Tyr Met Glu Gly Leu Phe 275 280 285 gac gag cgc tcc cgg ggc tac ttccga atc gac tct gcc acg ggc gcc 912 Asp Glu Arg Ser Arg Gly Tyr Phe ArgIle Asp Ser Ala Thr Gly Ala 290 295 300 gtg agc acg gac agc gta ctg gaccgc gag acc aag gag acg cac gtc 960 Val Ser Thr Asp Ser Val Leu Asp ArgGlu Thr Lys Glu Thr His Val 305 310 315 320 ctc agg gtg aaa gcc gtg gactac agt acg ccg ccg cgc tcg gcc acc 1008 Leu Arg Val Lys Ala Val Asp TyrSer Thr Pro Pro Arg Ser Ala Thr 325 330 335 acc tac atc act gtc ttg gtcaaa gac acc aac gac cac agc ccg gtc 1056 Thr Tyr Ile Thr Val Leu Val LysAsp Thr Asn Asp His Ser Pro Val 340 345 350 ttc gag cag tcg gag tac cgcgag cgc gtg cgg gag aac ctg gag gtg 1104 Phe Glu Gln Ser Glu Tyr Arg GluArg Val Arg Glu Asn Leu Glu Val 355 360 365 ggc tac gag gtg ctg acc atccgc gcc agc gac cgc gac tcg ccc atc 1152 Gly Tyr Glu Val Leu Thr Ile ArgAla Ser Asp Arg Asp Ser Pro Ile 370 375 380 aac gcc aac ttg cgt tac cgcgtg ttg ggg ggc gcg tgg gac gtc ttc 1200 Asn Ala Asn Leu Arg Tyr Arg ValLeu Gly Gly Ala Trp Asp Val Phe 385 390 395 400 cag ctc aac gag agc tctggc gtg gtg agc aca cgg gcg gtg ctg gac 1248 Gln Leu Asn Glu Ser Ser GlyVal Val Ser Thr Arg Ala Val Leu Asp 405 410 415 cgg gag gag gcg gcc gagtac cag ctc ctg gtg gag gcc aac gac cag 1296 Arg Glu Glu Ala Ala Glu TyrGln Leu Leu Val Glu Ala Asn Asp Gln 420 425 430 ggg cgc aat ccg ggc ccgctc agt gcc acg gcc acc gtg tac atc gag 1344 Gly Arg Asn Pro Gly Pro LeuSer Ala Thr Ala Thr Val Tyr Ile Glu 435 440 445 gtg gag gac gag aac gacaac tac ccc cag ttc agc gag cag aac tac 1392 Val Glu Asp Glu Asn Asp AsnTyr Pro Gln Phe Ser Glu Gln Asn Tyr 450 455 460 gtg gtc cag gtg ccc gaggac gtg ggg ctc aac acg gct gtg ctg cga 1440 Val Val Gln Val Pro Glu AspVal Gly Leu Asn Thr Ala Val Leu Arg 465 470 475 480 gtg cag gcc acg gaccgg gac cag ggc cag aac gcg gcc att cac tac 1488 Val Gln Ala Thr Asp ArgAsp Gln Gly Gln Asn Ala Ala Ile His Tyr 485 490 495 agc atc ctc agc gggaac gtg gcc ggc cag ttc tac ctg cac tcg ctg 1536 Ser Ile Leu Ser Gly AsnVal Ala Gly Gln Phe Tyr Leu His Ser Leu 500 505 510 agc ggg atc ctg gatgtg atc aac ccc ttg gat ttc gag gat gtc cag 1584 Ser Gly Ile Leu Asp ValIle Asn Pro Leu Asp Phe Glu Asp Val Gln 515 520 525 aaa tac tcg ctg agcatt aag gcc cag gat ggg ggc cgg ccc ccg ctc 1632 Lys Tyr Ser Leu Ser IleLys Ala Gln Asp Gly Gly Arg Pro Pro Leu 530 535 540 atc aat tct tca ggggtg gtg tct gtg cag gtg ctg gat gtc aac gac 1680 Ile Asn Ser Ser Gly ValVal Ser Val Gln Val Leu Asp Val Asn Asp 545 550 555 560 aac gag cct atcttt gtg agc agc ccc ttc cag gcc acg gtg ctg gag 1728 Asn Glu Pro Ile PheVal Ser Ser Pro Phe Gln Ala Thr Val Leu Glu 565 570 575 aat gtg ccc ctgggc tac ccc gtg gtg cac att cag gcg gtg gac gcg 1776 Asn Val Pro Leu GlyTyr Pro Val Val His Ile Gln Ala Val Asp Ala 580 585 590 gac tct gga gagaac gcc cgg ctg cac tat cgc ctg gtg gac acg gcc 1824 Asp Ser Gly Glu AsnAla Arg Leu His Tyr Arg Leu Val Asp Thr Ala 595 600 605 tcc acc ttt ctgggg ggc ggc agc gct ggg cct aag aat cct gcc ccc 1872 Ser Thr Phe Leu GlyGly Gly Ser Ala Gly Pro Lys Asn Pro Ala Pro 610 615 620 acc cct gac ttcccc ttc cag atc cac aac agc tcc ggt tgg atc aca 1920 Thr Pro Asp Phe ProPhe Gln Ile His Asn Ser Ser Gly Trp Ile Thr 625 630 635 640 gtg tgt gccgag ctg gac cgc gag gag gtg gag cac tac agc ttc ggg 1968 Val Cys Ala GluLeu Asp Arg Glu Glu Val Glu His Tyr Ser Phe Gly 645 650 655 gtg gag gcggtg gac cac ggc tcg ccc ccc atg agc tcc tcc acc agc 2016 Val Glu Ala ValAsp His Gly Ser Pro Pro Met Ser Ser Ser Thr Ser 660 665 670 gtg tcc atcacg gtg ctg gac gtg aat gac aac gac ccg gtg ttc acg 2064 Val Ser Ile ThrVal Leu Asp Val Asn Asp Asn Asp Pro Val Phe Thr 675 680 685 cag ccc acctac gag ctt cgt ctg aat gag gat gcg gcc gtg ggg agc 2112 Gln Pro Thr TyrGlu Leu Arg Leu Asn Glu Asp Ala Ala Val Gly Ser 690 695 700 agc gtg ctgacc ctg cag gcc cgc gac cgt gac gcc aac agt gtg att 2160 Ser Val Leu ThrLeu Gln Ala Arg Asp Arg Asp Ala Asn Ser Val Ile 705 710 715 720 acc taccag ctc aca ggc ggc aac acc cgg aac cgc ttt gca ctc agc 2208 Thr Tyr GlnLeu Thr Gly Gly Asn Thr Arg Asn Arg Phe Ala Leu Ser 725 730 735 agc cagaga ggg ggc ggc ctc atc acc ctg gcg cta cct ctg gac tac 2256 Ser Gln ArgGly Gly Gly Leu Ile Thr Leu Ala Leu Pro Leu Asp Tyr 740 745 750 aag caggag cag cag tac gtg ctg gcg gtg aca gca tcc gac ggc aca 2304 Lys Gln GluGln Gln Tyr Val Leu Ala Val Thr Ala Ser Asp Gly Thr 755 760 765 cgg tcgcac act gcg cat gtc cta atc aac gtc act gat gcc aac acc 2352 Arg Ser HisThr Ala His Val Leu Ile Asn Val Thr Asp Ala Asn Thr 770 775 780 cac aggcct gtc ttt cag agc tcc cat tac aca gtg agt gtc agt gag 2400 His Arg ProVal Phe Gln Ser Ser His Tyr Thr Val Ser Val Ser Glu 785 790 795 800 gacagg cct gtg ggc acc tcc att gct acc ctc agt gcc aac gat gag 2448 Asp ArgPro Val Gly Thr Ser Ile Ala Thr Leu Ser Ala Asn Asp Glu 805 810 815 gacaca gga gag aat gcc cgc atc acc tac gtg att cag gac ccc gtg 2496 Asp ThrGly Glu Asn Ala Arg Ile Thr Tyr Val Ile Gln Asp Pro Val 820 825 830 ccgcag ttc cgc att gac ccc gac agt ggc acc atg tac acc atg atg 2544 Pro GlnPhe Arg Ile Asp Pro Asp Ser Gly Thr Met Tyr Thr Met Met 835 840 845 gagctg gac tat gag aac cag gtc gcc tac acg ctg acc atc atg gcc 2592 Glu LeuAsp Tyr Glu Asn Gln Val Ala Tyr Thr Leu Thr Ile Met Ala 850 855 860 caggac aac ggc atc ccg cag aaa tca gac acc acc acc cta gag atc 2640 Gln AspAsn Gly Ile Pro Gln Lys Ser Asp Thr Thr Thr Leu Glu Ile 865 870 875 880ctc atc ctc gat gcc aat gac aat gca ccc cag ttc ctg tgg gat ttc 2688 LeuIle Leu Asp Ala Asn Asp Asn Ala Pro Gln Phe Leu Trp Asp Phe 885 890 895tac cag ggt tcc atc ttt gag gat gct cca ccc tcg acc agc atc ctc 2736 TyrGln Gly Ser Ile Phe Glu Asp Ala Pro Pro Ser Thr Ser Ile Leu 900 905 910cag gtc tct gcc acg gac cgg gac tca ggt ccc aat ggg cgt ctg ctg 2784 GlnVal Ser Ala Thr Asp Arg Asp Ser Gly Pro Asn Gly Arg Leu Leu 915 920 925tac acc ttc cag ggt ggg gac gac ggc gat ggg gac ttc tac atc gag 2832 TyrThr Phe Gln Gly Gly Asp Asp Gly Asp Gly Asp Phe Tyr Ile Glu 930 935 940ccc acg tcc ggt gtg att cgc acc cag cgc cgg ctg gac cgg gag aat 2880 ProThr Ser Gly Val Ile Arg Thr Gln Arg Arg Leu Asp Arg Glu Asn 945 950 955960 gtg gcc gtg tac aac ctt tgg gct ctg gct gtg gat cgg ggc agt ccc 2928Val Ala Val Tyr Asn Leu Trp Ala Leu Ala Val Asp Arg Gly Ser Pro 965 970975 act ccc ctt agc gcc tcg gta gaa atc cag gtg acc atc ttg gac att 2976Thr Pro Leu Ser Ala Ser Val Glu Ile Gln Val Thr Ile Leu Asp Ile 980 985990 aat gac aat gcc ccc atg ttt gag aag gac gaa ctg gag ctg ttt gtt 3024Asn Asp Asn Ala Pro Met Phe Glu Lys Asp Glu Leu Glu Leu Phe Val 995 10001005 gag gag aac aac cca gtg ggg tcg gtg gtg gca aag att cgt gct aac3072 Glu Glu Asn Asn Pro Val Gly Ser Val Val Ala Lys Ile Arg Ala Asn1010 1015 1020 gac cct gat gaa ggc cct aat gcc cag atc atg tat cag attgtg gaa 3120 Asp Pro Asp Glu Gly Pro Asn Ala Gln Ile Met Tyr Gln Ile ValGlu 1025 1030 1035 1040 ggg gac atg cgg cat ttc ttc cag ctg gac ctg ctcaac ggg gac ctg 3168 Gly Asp Met Arg His Phe Phe Gln Leu Asp Leu Leu AsnGly Asp Leu 1045 1050 1055 cgt gcc atg gtg gag ctg gac ttt gag gtc cggcgg gag tat gtg ctg 3216 Arg Ala Met Val Glu Leu Asp Phe Glu Val Arg ArgGlu Tyr Val Leu 1060 1065 1070 gtg gtg cag gcc acg tcg gct ccg ctg gtgagc cga gcc acg gtg cac 3264 Val Val Gln Ala Thr Ser Ala Pro Leu Val SerArg Ala Thr Val His 1075 1080 1085 atc ctt ctc gtg gac cag aat gac aacccg cct gtg ctg ccc gac ttc 3312 Ile Leu Leu Val Asp Gln Asn Asp Asn ProPro Val Leu Pro Asp Phe 1090 1095 1100 cag atc ctc ttc aac aac tat gtcacc aac aag tcc aac agt ttc ccc 3360 Gln Ile Leu Phe Asn Asn Tyr Val ThrAsn Lys Ser Asn Ser Phe Pro 1105 1110 1115 1120 acc ggc gtg atc ggc tgcatc ccg gcc cat gac ccc gac gtg tca gac 3408 Thr Gly Val Ile Gly Cys IlePro Ala His Asp Pro Asp Val Ser Asp 1125 1130 1135 agc ctc aac tac accttc gtg cag ggc aac gag ctg cgc ctg ttg ctg 3456 Ser Leu Asn Tyr Thr PheVal Gln Gly Asn Glu Leu Arg Leu Leu Leu 1140 1145 1150 ctg gac ccc gccacg ggc gaa ctg cag ctc agc cgc gac ctg gac aac 3504 Leu Asp Pro Ala ThrGly Glu Leu Gln Leu Ser Arg Asp Leu Asp Asn 1155 1160 1165 aac cgg ccgctg gag gcg ctc atg gag gtg tct gtg tct gat ggc atc 3552 Asn Arg Pro LeuGlu Ala Leu Met Glu Val Ser Val Ser Asp Gly Ile 1170 1175 1180 cac agcgtc acg gcc ttc tgc acc ctg cgt gtc acc atc atc acg gac 3600 His Ser ValThr Ala Phe Cys Thr Leu Arg Val Thr Ile Ile Thr Asp 1185 1190 1195 1200gac atg ctg acc aac agc atc act gtc cgc ctg gag aac atg tcc cag 3648 AspMet Leu Thr Asn Ser Ile Thr Val Arg Leu Glu Asn Met Ser Gln 1205 12101215 gag aag ttc ctg tcc ccg ctg ctg gcc ctc ttc gtg gag ggg gtg gcc3696 Glu Lys Phe Leu Ser Pro Leu Leu Ala Leu Phe Val Glu Gly Val Ala1220 1225 1230 gcc gtg ctg tcc acc acc aag gac gac gtc ttc gtc ttc aacgtc cag 3744 Ala Val Leu Ser Thr Thr Lys Asp Asp Val Phe Val Phe Asn ValGln 1235 1240 1245 aac gac acc gac gtc agc tcc aac atc ctg aac gtg accttc tcg gcg 3792 Asn Asp Thr Asp Val Ser Ser Asn Ile Leu Asn Val Thr PheSer Ala 1250 1255 1260 ctg ctg cct ggc ggc gtc cgc ggc cag ttc ttc ccgtcg gag gac ctg 3840 Leu Leu Pro Gly Gly Val Arg Gly Gln Phe Phe Pro SerGlu Asp Leu 1265 1270 1275 1280 cag gag cag atc tac ctg aat cgg acg ctgctg acc acc atc tcc acg 3888 Gln Glu Gln Ile Tyr Leu Asn Arg Thr Leu LeuThr Thr Ile Ser Thr 1285 1290 1295 cag cgc gtg ctg ccc ttc gac gac aacatc tgc ctg cgc gag ccc tgc 3936 Gln Arg Val Leu Pro Phe Asp Asp Asn IleCys Leu Arg Glu Pro Cys 1300 1305 1310 gag aac tac atg aag tgc gtg tccgtt ctg cga ttc gac agc tcc gcg 3984 Glu Asn Tyr Met Lys Cys Val Ser ValLeu Arg Phe Asp Ser Ser Ala 1315 1320 1325 ccc ttc ctc agc tcc acc accgtg ctc ttc cgg ccc atc cac ccc atc 4032 Pro Phe Leu Ser Ser Thr Thr ValLeu Phe Arg Pro Ile His Pro Ile 1330 1335 1340 aac ggc ctg cgc tgc cgctgc ccg ccc ggc ttc acc ggc gac tac tgc 4080 Asn Gly Leu Arg Cys Arg CysPro Pro Gly Phe Thr Gly Asp Tyr Cys 1345 1350 1355 1360 gag acg gag atcgac ctc tgc tac tcc gac ccg tgc ggc gcc aac ggc 4128 Glu Thr Glu Ile AspLeu Cys Tyr Ser Asp Pro Cys Gly Ala Asn Gly 1365 1370 1375 cgc tgc cgcagc cgc gag ggc ggc tac acc tgc gag tgc ttc gag gac 4176 Arg Cys Arg SerArg Glu Gly Gly Tyr Thr Cys Glu Cys Phe Glu Asp 1380 1385 1390 ttc actgga gag cac tgt gag gtg gat gcc cgc tca ggc cgc tgt gcc 4224 Phe Thr GlyGlu His Cys Glu Val Asp Ala Arg Ser Gly Arg Cys Ala 1395 1400 1405 aacggg gtg tgc aag aac ggg ggc acc tgc gtg aac ctg ctc atc ggc 4272 Asn GlyVal Cys Lys Asn Gly Gly Thr Cys Val Asn Leu Leu Ile Gly 1410 1415 1420ggc ttc cac tgc gtg tgt cct cct ggc gag tat gag agg ccc tac tgt 4320 GlyPhe His Cys Val Cys Pro Pro Gly Glu Tyr Glu Arg Pro Tyr Cys 1425 14301435 1440 gag gtg acc acc agg agc ttc ccg ccc cag tcc ttc gtc acc ttccgg 4368 Glu Val Thr Thr Arg Ser Phe Pro Pro Gln Ser Phe Val Thr Phe Arg1445 1450 1455 ggc ctg aga cag cgc ttc cac ttc acc atc tcc ctc acg tttgcc act 4416 Gly Leu Arg Gln Arg Phe His Phe Thr Ile Ser Leu Thr Phe AlaThr 1460 1465 1470 cag gaa agg aac ggc ttg ctt ctc tac aac ggc cgc ttcaat gag aag 4464 Gln Glu Arg Asn Gly Leu Leu Leu Tyr Asn Gly Arg Phe AsnGlu Lys 1475 1480 1485 cac gac ttc atc gcc ctg gag atc gtg gac gag caggtg cag ctc acc 4512 His Asp Phe Ile Ala Leu Glu Ile Val Asp Glu Gln ValGln Leu Thr 1490 1495 1500 ttc tct gca ggc gag aca aca acg acc gtg gcaccg aag gtt ccc agt 4560 Phe Ser Ala Gly Glu Thr Thr Thr Thr Val Ala ProLys Val Pro Ser 1505 1510 1515 1520 ggt gtg agt gac ggg cgg tgg cac tctgtg cag gtg cag tac tac aac 4608 Gly Val Ser Asp Gly Arg Trp His Ser ValGln Val Gln Tyr Tyr Asn 1525 1530 1535 aag ccc aat att ggc cac ctg ggcctg ccc cat ggg ccg tcc ggg gaa 4656 Lys Pro Asn Ile Gly His Leu Gly LeuPro His Gly Pro Ser Gly Glu 1540 1545 1550 aag atg gcc gtg gtg aca gtggat gat tgt gac aca acc atg gct gtg 4704 Lys Met Ala Val Val Thr Val AspAsp Cys Asp Thr Thr Met Ala Val 1555 1560 1565 cgc ttt gga aag gac atcggg aac tac agc tgc gct gcc cag ggc act 4752 Arg Phe Gly Lys Asp Ile GlyAsn Tyr Ser Cys Ala Ala Gln Gly Thr 1570 1575 1580 cag acc ggc tcc aagaag tcc ctg gat ctg acc ggc cct cta ctc ctg 4800 Gln Thr Gly Ser Lys LysSer Leu Asp Leu Thr Gly Pro Leu Leu Leu 1585 1590 1595 1600 ggg ggt gtcccc aac ctg cca gaa gac ttc cca gtg cac aac cgg cag 4848 Gly Gly Val ProAsn Leu Pro Glu Asp Phe Pro Val His Asn Arg Gln 1605 1610 1615 ttc gtgggc tgc atg cgg aac ctg tca gtc gac ggc aaa aat gtg gac 4896 Phe Val GlyCys Met Arg Asn Leu Ser Val Asp Gly Lys Asn Val Asp 1620 1625 1630 atggcc gga ttc atc gcc aac aat ggc acc cgg gaa ggc tgc gct gct 4944 Met AlaGly Phe Ile Ala Asn Asn Gly Thr Arg Glu Gly Cys Ala Ala 1635 1640 1645cgg agg aac ttc tgc gat ggg agg cgg tgt cag aat gga ggc acc tgt 4992 ArgArg Asn Phe Cys Asp Gly Arg Arg Cys Gln Asn Gly Gly Thr Cys 1650 16551660 gtc aac agg tgg aat atg tat ctg tgt gag tgt cca ctc cga ttc ggc5040 Val Asn Arg Trp Asn Met Tyr Leu Cys Glu Cys Pro Leu Arg Phe Gly1665 1670 1675 1680 ggg aag aac tgt gag caa gcc atg cct cac ccc cag ctcttc agc ggt 5088 Gly Lys Asn Cys Glu Gln Ala Met Pro His Pro Gln Leu PheSer Gly 1685 1690 1695 gag agc gtc gtg tcc tgg agt gac ctg aac atc atcatc tct gtg ccc 5136 Glu Ser Val Val Ser Trp Ser Asp Leu Asn Ile Ile IleSer Val Pro 1700 1705 1710 tgg tac ctg ggg ctc atg ttc cgg acc cgg aaggag gac agc gtt ctg 5184 Trp Tyr Leu Gly Leu Met Phe Arg Thr Arg Lys GluAsp Ser Val Leu 1715 1720 1725 atg gag gcc acc agt ggt ggg ccc acc agcttt cgc ctc cag atc ctg 5232 Met Glu Ala Thr Ser Gly Gly Pro Thr Ser PheArg Leu Gln Ile Leu 1730 1735 1740 aac aac tac ctc cag ttt gag gtg tcccac ggc ccc tcc gat gtg gag 5280 Asn Asn Tyr Leu Gln Phe Glu Val Ser HisGly Pro Ser Asp Val Glu 1745 1750 1755 1760 tcc gtg atg ctg tcc ggg ttgcgg gtg acc gac ggg gag tgg cac cac 5328 Ser Val Met Leu Ser Gly Leu ArgVal Thr Asp Gly Glu Trp His His 1765 1770 1775 ctg ctg atc gag ctg aagaat gtt aag gag gac agt gag atg aag cac 5376 Leu Leu Ile Glu Leu Lys AsnVal Lys Glu Asp Ser Glu Met Lys His 1780 1785 1790 ctg gtc acc atg accttg gac tat ggg atg gac cag aac aag gca gat 5424 Leu Val Thr Met Thr LeuAsp Tyr Gly Met Asp Gln Asn Lys Ala Asp 1795 1800 1805 atc ggg ggc atgctt ccc ggg ctg acg gta agg agc gtg gtg gtc gga 5472 Ile Gly Gly Met LeuPro Gly Leu Thr Val Arg Ser Val Val Val Gly 1810 1815 1820 ggc gcc tctgaa gac aag gtc tcc gtg cgc cgt gga ttc cga ggc tgc 5520 Gly Ala Ser GluAsp Lys Val Ser Val Arg Arg Gly Phe Arg Gly Cys 1825 1830 1835 1840 atgcag gga gtg agg atg ggg ggg acg ccc acc aac gtc gcc acc ctg 5568 Met GlnGly Val Arg Met Gly Gly Thr Pro Thr Asn Val Ala Thr Leu 1845 1850 1855aac atg aac aac gca ctc aag gtc agg gtg aag gac ggc tgt gat gtg 5616 AsnMet Asn Asn Ala Leu Lys Val Arg Val Lys Asp Gly Cys Asp Val 1860 18651870 gac gac ccc tgt acc tcg agc ccc tgt ccc ccc aat agc cgc tgc cac5664 Asp Asp Pro Cys Thr Ser Ser Pro Cys Pro Pro Asn Ser Arg Cys His1875 1880 1885 gac gcc tgg gag gac tac agc tgc gtc tgt gac aaa ggg tacctt gga 5712 Asp Ala Trp Glu Asp Tyr Ser Cys Val Cys Asp Lys Gly Tyr LeuGly 1890 1895 1900 ata aac tgt gtg gat gcc tgt cac ctg aac ccc tgc gagaac atg ggg 5760 Ile Asn Cys Val Asp Ala Cys His Leu Asn Pro Cys Glu AsnMet Gly 1905 1910 1915 1920 gcc tgc gtg cgc tcc ccc ggc tcc ccg cag ggctac gtg tgc gag tgt 5808 Ala Cys Val Arg Ser Pro Gly Ser Pro Gln Gly TyrVal Cys Glu Cys 1925 1930 1935 ggg ccc agt cac tac ggg ccg tac tgt gagaac aaa ctc gac ctt ccg 5856 Gly Pro Ser His Tyr Gly Pro Tyr Cys Glu AsnLys Leu Asp Leu Pro 1940 1945 1950 tgc ccc aga ggc tgg tgg ggg aac cccgtc tgt gga ccc tgc cac tgt 5904 Cys Pro Arg Gly Trp Trp Gly Asn Pro ValCys Gly Pro Cys His Cys 1955 1960 1965 gcc gtc agc aaa ggc ttt gat cccgac tgt aat aag acc aac ggc cag 5952 Ala Val Ser Lys Gly Phe Asp Pro AspCys Asn Lys Thr Asn Gly Gln 1970 1975 1980 tgc caa tgc aag gag aat tactac aag ctc cta gcc cag gac acc tgt 6000 Cys Gln Cys Lys Glu Asn Tyr TyrLys Leu Leu Ala Gln Asp Thr Cys 1985 1990 1995 2000 ctg ccc tgc gac tgcttc ccc cat ggc tcc cac agc cgc act tgc gac 6048 Leu Pro Cys Asp Cys PhePro His Gly Ser His Ser Arg Thr Cys Asp 2005 2010 2015 atg gcc acc gggcag tgt gcc tgc aag ccc ggc gtc atc ggc cgc cag 6096 Met Ala Thr Gly GlnCys Ala Cys Lys Pro Gly Val Ile Gly Arg Gln 2020 2025 2030 tgc aac cgctgc gac aac ccg ttt gcc gag gtc acc acg ctc ggc tgt 6144 Cys Asn Arg CysAsp Asn Pro Phe Ala Glu Val Thr Thr Leu Gly Cys 2035 2040 2045 gaa gtgatc tac aat ggc tgt ccc aaa gca ttt gag gcc ggc atc tgg 6192 Glu Val IleTyr Asn Gly Cys Pro Lys Ala Phe Glu Ala Gly Ile Trp 2050 2055 2060 tggcca cag acc aag ttc ggg cag ccg gct gcg gtg cca tgc cct aag 6240 Trp ProGln Thr Lys Phe Gly Gln Pro Ala Ala Val Pro Cys Pro Lys 2065 2070 20752080 gga tcc gtt gga aat gcg gtc cga cac tgc agc ggg gag aag ggc tgg6288 Gly Ser Val Gly Asn Ala Val Arg His Cys Ser Gly Glu Lys Gly Trp2085 2090 2095 ctg ccc cca gag ctc ttt aac tgt acc acc atc tcc ttc gtggac ctc 6336 Leu Pro Pro Glu Leu Phe Asn Cys Thr Thr Ile Ser Phe Val AspLeu 2100 2105 2110 agg gcc atg aat gag aag ctg agc cgc aat gag acg caggtg gac ggc 6384 Arg Ala Met Asn Glu Lys Leu Ser Arg Asn Glu Thr Gln ValAsp Gly 2115 2120 2125 gcc agg gcc ctg cag ctg gtg agg gcg ctg cgc agtgct aca cag cac 6432 Ala Arg Ala Leu Gln Leu Val Arg Ala Leu Arg Ser AlaThr Gln His 2130 2135 2140 acg ggc acg ctc ttt ggc aat gac gtg cgc acggcc tac cag ctg ctg 6480 Thr Gly Thr Leu Phe Gly Asn Asp Val Arg Thr AlaTyr Gln Leu Leu 2145 2150 2155 2160 ggc cac gtc ctt cag cac gag agc tggcag cag ggc ttc gac ctg gca 6528 Gly His Val Leu Gln His Glu Ser Trp GlnGln Gly Phe Asp Leu Ala 2165 2170 2175 gcc acg cag gac gcc gac ttt cacgag gac gtc atc cac tcg ggc agc 6576 Ala Thr Gln Asp Ala Asp Phe His GluAsp Val Ile His Ser Gly Ser 2180 2185 2190 gcc ctc ctg gcc cca gcc accagg gcg gcg tgg gag cag atc cag cgg 6624 Ala Leu Leu Ala Pro Ala Thr ArgAla Ala Trp Glu Gln Ile Gln Arg 2195 2200 2205 agc gag ggc ggc acg gcacag ctg ctc cgg cgc ctc gag ggc tac ttc 6672 Ser Glu Gly Gly Thr Ala GlnLeu Leu Arg Arg Leu Glu Gly Tyr Phe 2210 2215 2220 agc aac gtg gca cgcaac gtg cgg cgg acg tac ctg cgg ccc ttc gtc 6720 Ser Asn Val Ala Arg AsnVal Arg Arg Thr Tyr Leu Arg Pro Phe Val 2225 2230 2235 2240 atc gtc accgcc aac atg att ctt gct gtc gac atc ttt gac aag ttc 6768 Ile Val Thr AlaAsn Met Ile Leu Ala Val Asp Ile Phe Asp Lys Phe 2245 2250 2255 aac tttacg gga gcc agg gtc ccg cga ttc gac acc atc cat gaa gag 6816 Asn Phe ThrGly Ala Arg Val Pro Arg Phe Asp Thr Ile His Glu Glu 2260 2265 2270 ttcccc agg gag ctg gag tcc tcc gtc tcc ttc cca gcc gac ttc ttc 6864 Phe ProArg Glu Leu Glu Ser Ser Val Ser Phe Pro Ala Asp Phe Phe 2275 2280 2285aga cca cct gaa gaa aaa gaa ggc ccc ctg ctg agg ccg gct ggc cgg 6912 ArgPro Pro Glu Glu Lys Glu Gly Pro Leu Leu Arg Pro Ala Gly Arg 2290 22952300 agg acc acc ccg cag acc acg cgc ccg ggg cct ggc acc gag agg gag6960 Arg Thr Thr Pro Gln Thr Thr Arg Pro Gly Pro Gly Thr Glu Arg Glu2305 2310 2315 2320 gcc ccg atc agc agg cgg agg cga cac cct gat gac gctggc cag ttc 7008 Ala Pro Ile Ser Arg Arg Arg Arg His Pro Asp Asp Ala GlyGln Phe 2325 2330 2335 gcc gtc gct ctg gtc atc att tac cgc acc ctg gggcag ctc ctg ccc 7056 Ala Val Ala Leu Val Ile Ile Tyr Arg Thr Leu Gly GlnLeu Leu Pro 2340 2345 2350 gag cgc tac gac ccc gac cgt cgc agc ctc cggttg cct cac cgg ccc 7104 Glu Arg Tyr Asp Pro Asp Arg Arg Ser Leu Arg LeuPro His Arg Pro 2355 2360 2365 atc att aat acc ccg atg gtg agc acg ctggtg tac agc gag ggg gct 7152 Ile Ile Asn Thr Pro Met Val Ser Thr Leu ValTyr Ser Glu Gly Ala 2370 2375 2380 ccg ctc ccg aga ccc ctg gag agg cccgtc ctg gtg gag ttc gcc ctg 7200 Pro Leu Pro Arg Pro Leu Glu Arg Pro ValLeu Val Glu Phe Ala Leu 2385 2390 2395 2400 ctg gag gtg gag gag cga accaag cct gtc tgc gtg ttc tgg aac cac 7248 Leu Glu Val Glu Glu Arg Thr LysPro Val Cys Val Phe Trp Asn His 2405 2410 2415 tcc ctg gcc gtt ggt gggacg gga ggg tgg tct gcc cgg ggc tgc gag 7296 Ser Leu Ala Val Gly Gly ThrGly Gly Trp Ser Ala Arg Gly Cys Glu 2420 2425 2430 ctc ctg tcc agg aaccgg aca cat gtc gcc tgc cag tgc agc cac aca 7344 Leu Leu Ser Arg Asn ArgThr His Val Ala Cys Gln Cys Ser His Thr 2435 2440 2445 gcc agc ttt gcggtg ctc atg gat atc tcc agg cgt gag aac ggg gag 7392 Ala Ser Phe Ala ValLeu Met Asp Ile Ser Arg Arg Glu Asn Gly Glu 2450 2455 2460 gtc ctg cctctg aag att gtc acc tat gcc gct gtg tcc ttg tca ctg 7440 Val Leu Pro LeuLys Ile Val Thr Tyr Ala Ala Val Ser Leu Ser Leu 2465 2470 2475 2480 gcagcc ctg ctg gtg gcc ttc gtc ctc ctg agc ctg gtc cgc atg ctg 7488 Ala AlaLeu Leu Val Ala Phe Val Leu Leu Ser Leu Val Arg Met Leu 2485 2490 2495cgc tcc aac ctg cac agc att cac aag cac ctc gcc gtg gcg ctc ttc 7536 ArgSer Asn Leu His Ser Ile His Lys His Leu Ala Val Ala Leu Phe 2500 25052510 ctc tct cag ctg gtg ttc gtg att ggg atc aac cag acg gaa aac ccg7584 Leu Ser Gln Leu Val Phe Val Ile Gly Ile Asn Gln Thr Glu Asn Pro2515 2520 2525 ttt ctg tgc aca gtg gtt gcc atc ctc ctc cac tac atc tacatg agc 7632 Phe Leu Cys Thr Val Val Ala Ile Leu Leu His Tyr Ile Tyr MetSer 2530 2535 2540 acc ttt gcc tgg acc ctc gtg gag agc ctg cat gtc taccgc atg ctg 7680 Thr Phe Ala Trp Thr Leu Val Glu Ser Leu His Val Tyr ArgMet Leu 2545 2550 2555 2560 acc gag gtg cgc aac atc gac acg ggg ccc atgcgg ttc tac tac gtc 7728 Thr Glu Val Arg Asn Ile Asp Thr Gly Pro Met ArgPhe Tyr Tyr Val 2565 2570 2575 gtg ggc tgg ggc atc ccg gcc att gtc acagga ctg gcg gtc ggc ctg 7776 Val Gly Trp Gly Ile Pro Ala Ile Val Thr GlyLeu Ala Val Gly Leu 2580 2585 2590 gac ccc cag ggc tac ggg aac ccc gacttc tgc tgg ctg tcg ctt caa 7824 Asp Pro Gln Gly Tyr Gly Asn Pro Asp PheCys Trp Leu Ser Leu Gln 2595 2600 2605 gac acc ctg att tgg agc ttt gcgggg ccc atc gga gct gtt ata atc 7872 Asp Thr Leu Ile Trp Ser Phe Ala GlyPro Ile Gly Ala Val Ile Ile 2610 2615 2620 atc aac aca gtc act tct gtccta tct gca aag gtt tcc tgc caa aga 7920 Ile Asn Thr Val Thr Ser Val LeuSer Ala Lys Val Ser Cys Gln Arg 2625 2630 2635 2640 aag cac cat tat tatggg aaa aaa ggg atc gtc tcc ctg ctg agg acc 7968 Lys His His Tyr Tyr GlyLys Lys Gly Ile Val Ser Leu Leu Arg Thr 2645 2650 2655 gca ttc ctc ctgctg ctg ctc atc agc gcc acc tgg ctg ctg ggg ctg 8016 Ala Phe Leu Leu LeuLeu Leu Ile Ser Ala Thr Trp Leu Leu Gly Leu 2660 2665 2670 ctg gct gtgaac cgc gat gca ctg agc ttt cac tac ctc ttc gcc atc 8064 Leu Ala Val AsnArg Asp Ala Leu Ser Phe His Tyr Leu Phe Ala Ile 2675 2680 2685 ttc agcggc tta cag ggc ccc ttc gtc ctc ctt ttc cac tgc gtg ctc 8112 Phe Ser GlyLeu Gln Gly Pro Phe Val Leu Leu Phe His Cys Val Leu 2690 2695 2700 aaccag gag gtc cgg aag cac ctg aag ggc gtg ctc ggc ggg agg aag 8160 Asn GlnGlu Val Arg Lys His Leu Lys Gly Val Leu Gly Gly Arg Lys 2705 2710 27152720 ctg cac ctg gag gac tcc gcc acc acc agg gcc acc ctg ctg acg cgc8208 Leu His Leu Glu Asp Ser Ala Thr Thr Arg Ala Thr Leu Leu Thr Arg2725 2730 2735 tcc ctc aac tgc aac acc acc ttc ggt gac ggg cct gac atgctg cgc 8256 Ser Leu Asn Cys Asn Thr Thr Phe Gly Asp Gly Pro Asp Met LeuArg 2740 2745 2750 aca gac ttg ggc gag tcc acc gcc tcg ctg gac agc atcgtc agg gat 8304 Thr Asp Leu Gly Glu Ser Thr Ala Ser Leu Asp Ser Ile ValArg Asp 2755 2760 2765 gaa ggg atc cag aag ctc ggc gtg tcc tct ggg ctggtg agg ggc agc 8352 Glu Gly Ile Gln Lys Leu Gly Val Ser Ser Gly Leu ValArg Gly Ser 2770 2775 2780 cac gga gag cca gac gcg tcc ctc atg ccc aggagc tgc aag gat ccc 8400 His Gly Glu Pro Asp Ala Ser Leu Met Pro Arg SerCys Lys Asp Pro 2785 2790 2795 2800 cct ggc cac gat tcc gac tca gat agcgag ctg tcc ctg gat gag cag 8448 Pro Gly His Asp Ser Asp Ser Asp Ser GluLeu Ser Leu Asp Glu Gln 2805 2810 2815 agc agc tct tac gcc tcc tca cactcg tca gac agc gag gac gat ggg 8496 Ser Ser Ser Tyr Ala Ser Ser His SerSer Asp Ser Glu Asp Asp Gly 2820 2825 2830 gtg gga gct gag gaa aaa tgggac ccg gcc agg ggc gcc gtc cac agc 8544 Val Gly Ala Glu Glu Lys Trp AspPro Ala Arg Gly Ala Val His Ser 2835 2840 2845 acc ccc aaa ggg gac gctgtg gcc aac cac gtt ccg gcc ggc tgg ccc 8592 Thr Pro Lys Gly Asp Ala ValAla Asn His Val Pro Ala Gly Trp Pro 2850 2855 2860 gac cag agc ctg gctgag agt gac agt gag gac ccc agc ggc aag ccc 8640 Asp Gln Ser Leu Ala GluSer Asp Ser Glu Asp Pro Ser Gly Lys Pro 2865 2870 2875 2880 cgc ctg aaggtg gag acc aag gtc agc gtg gag ctg cac cgc gag gag 8688 Arg Leu Lys ValGlu Thr Lys Val Ser Val Glu Leu His Arg Glu Glu 2885 2890 2895 cag ggcagt cac cgt gga gag tac ccc ccg gac cag gag agc ggg ggc 8736 Gln Gly SerHis Arg Gly Glu Tyr Pro Pro Asp Gln Glu Ser Gly Gly 2900 2905 2910 gcagcc agg ctt gct agc agc cag ccc cca gag cag agg aaa ggc atc 8784 Ala AlaArg Leu Ala Ser Ser Gln Pro Pro Glu Gln Arg Lys Gly Ile 2915 2920 2925ttg aaa aat aaa gtc acc tac ccg ccg ccg ctg acg ctg acg gag cag 8832 LeuLys Asn Lys Val Thr Tyr Pro Pro Pro Leu Thr Leu Thr Glu Gln 2930 29352940 acg ctg aag ggc cgg ctc cgg gag aag ctg gcc gac tgt gag cag agc8880 Thr Leu Lys Gly Arg Leu Arg Glu Lys Leu Ala Asp Cys Glu Gln Ser2945 2950 2955 2960 ccc aca tcc tcg cgc acg tct tcc ctg ggc tct ggc ggcccc gac tgc 8928 Pro Thr Ser Ser Arg Thr Ser Ser Leu Gly Ser Gly Gly ProAsp Cys 2965 2970 2975 gcc atc aca gtc aag agc cct ggg agg gag ccg gggcgt gac cac ctc 8976 Ala Ile Thr Val Lys Ser Pro Gly Arg Glu Pro Gly ArgAsp His Leu 2980 2985 2990 aac ggg gtg gcc atg aat gtg cgc act ggg agcgcc cag gcc gat ggc 9024 Asn Gly Val Ala Met Asn Val Arg Thr Gly Ser AlaGln Ala Asp Gly 2995 3000 3005 tcc gac tct gag aaa ccg tga 9045 Ser AspSer Glu Lys Pro 3010 2 3014 PRT Homo sapiens 2 Met Ala Pro Pro Pro ProPro Val Leu Pro Val Leu Leu Leu Leu Ala 1 5 10 15 Ala Ala Ala Ala LeuPro Ala Met Gly Leu Arg Ala Ala Ala Trp Glu 20 25 30 Pro Arg Val Pro GlyGly Thr Arg Ala Phe Ala Leu Arg Pro Gly Cys 35 40 45 Thr Tyr Ala Val GlyAla Ala Cys Thr Pro Arg Ala Pro Arg Glu Leu 50 55 60 Leu Asp Val Gly ArgAsp Gly Arg Leu Ala Gly Arg Arg Arg Val Ser 65 70 75 80 Gly Ala Gly ArgPro Leu Pro Leu Gln Val Arg Leu Val Ala Arg Ser 85 90 95 Ala Pro Thr AlaLeu Ser Arg Arg Leu Arg Ala Arg Thr His Leu Pro 100 105 110 Gly Cys GlyAla Arg Ala Arg Leu Cys Gly Thr Gly Ala Arg Leu Cys 115 120 125 Gly AlaLeu Cys Phe Pro Val Pro Gly Gly Cys Ala Ala Ala Gln His 130 135 140 SerAla Leu Ala Ala Pro Thr Thr Leu Pro Ala Cys Arg Cys Pro Pro 145 150 155160 Arg Pro Arg Pro Arg Cys Pro Gly Arg Pro Ile Cys Leu Pro Pro Gly 165170 175 Gly Ser Val Arg Leu Arg Leu Leu Cys Ala Leu Arg Arg Ala Ala Gly180 185 190 Ala Val Arg Val Gly Leu Ala Leu Glu Ala Ala Thr Ala Gly ThrPro 195 200 205 Ser Ala Ser Pro Ser Pro Ser Pro Pro Leu Pro Pro Asn LeuPro Glu 210 215 220 Ala Arg Ala Gly Pro Ala Arg Arg Ala Arg Arg Gly ThrSer Gly Arg 225 230 235 240 Gly Ser Leu Lys Phe Pro Met Pro Asn Tyr GlnVal Ala Leu Phe Glu 245 250 255 Asn Glu Pro Ala Gly Thr Leu Ile Leu GlnLeu His Ala His Tyr Thr 260 265 270 Ile Glu Gly Glu Glu Glu Arg Val SerTyr Tyr Met Glu Gly Leu Phe 275 280 285 Asp Glu Arg Ser Arg Gly Tyr PheArg Ile Asp Ser Ala Thr Gly Ala 290 295 300 Val Ser Thr Asp Ser Val LeuAsp Arg Glu Thr Lys Glu Thr His Val 305 310 315 320 Leu Arg Val Lys AlaVal Asp Tyr Ser Thr Pro Pro Arg Ser Ala Thr 325 330 335 Thr Tyr Ile ThrVal Leu Val Lys Asp Thr Asn Asp His Ser Pro Val 340 345 350 Phe Glu GlnSer Glu Tyr Arg Glu Arg Val Arg Glu Asn Leu Glu Val 355 360 365 Gly TyrGlu Val Leu Thr Ile Arg Ala Ser Asp Arg Asp Ser Pro Ile 370 375 380 AsnAla Asn Leu Arg Tyr Arg Val Leu Gly Gly Ala Trp Asp Val Phe 385 390 395400 Gln Leu Asn Glu Ser Ser Gly Val Val Ser Thr Arg Ala Val Leu Asp 405410 415 Arg Glu Glu Ala Ala Glu Tyr Gln Leu Leu Val Glu Ala Asn Asp Gln420 425 430 Gly Arg Asn Pro Gly Pro Leu Ser Ala Thr Ala Thr Val Tyr IleGlu 435 440 445 Val Glu Asp Glu Asn Asp Asn Tyr Pro Gln Phe Ser Glu GlnAsn Tyr 450 455 460 Val Val Gln Val Pro Glu Asp Val Gly Leu Asn Thr AlaVal Leu Arg 465 470 475 480 Val Gln Ala Thr Asp Arg Asp Gln Gly Gln AsnAla Ala Ile His Tyr 485 490 495 Ser Ile Leu Ser Gly Asn Val Ala Gly GlnPhe Tyr Leu His Ser Leu 500 505 510 Ser Gly Ile Leu Asp Val Ile Asn ProLeu Asp Phe Glu Asp Val Gln 515 520 525 Lys Tyr Ser Leu Ser Ile Lys AlaGln Asp Gly Gly Arg Pro Pro Leu 530 535 540 Ile Asn Ser Ser Gly Val ValSer Val Gln Val Leu Asp Val Asn Asp 545 550 555 560 Asn Glu Pro Ile PheVal Ser Ser Pro Phe Gln Ala Thr Val Leu Glu 565 570 575 Asn Val Pro LeuGly Tyr Pro Val Val His Ile Gln Ala Val Asp Ala 580 585 590 Asp Ser GlyGlu Asn Ala Arg Leu His Tyr Arg Leu Val Asp Thr Ala 595 600 605 Ser ThrPhe Leu Gly Gly Gly Ser Ala Gly Pro Lys Asn Pro Ala Pro 610 615 620 ThrPro Asp Phe Pro Phe Gln Ile His Asn Ser Ser Gly Trp Ile Thr 625 630 635640 Val Cys Ala Glu Leu Asp Arg Glu Glu Val Glu His Tyr Ser Phe Gly 645650 655 Val Glu Ala Val Asp His Gly Ser Pro Pro Met Ser Ser Ser Thr Ser660 665 670 Val Ser Ile Thr Val Leu Asp Val Asn Asp Asn Asp Pro Val PheThr 675 680 685 Gln Pro Thr Tyr Glu Leu Arg Leu Asn Glu Asp Ala Ala ValGly Ser 690 695 700 Ser Val Leu Thr Leu Gln Ala Arg Asp Arg Asp Ala AsnSer Val Ile 705 710 715 720 Thr Tyr Gln Leu Thr Gly Gly Asn Thr Arg AsnArg Phe Ala Leu Ser 725 730 735 Ser Gln Arg Gly Gly Gly Leu Ile Thr LeuAla Leu Pro Leu Asp Tyr 740 745 750 Lys Gln Glu Gln Gln Tyr Val Leu AlaVal Thr Ala Ser Asp Gly Thr 755 760 765 Arg Ser His Thr Ala His Val LeuIle Asn Val Thr Asp Ala Asn Thr 770 775 780 His Arg Pro Val Phe Gln SerSer His Tyr Thr Val Ser Val Ser Glu 785 790 795 800 Asp Arg Pro Val GlyThr Ser Ile Ala Thr Leu Ser Ala Asn Asp Glu 805 810 815 Asp Thr Gly GluAsn Ala Arg Ile Thr Tyr Val Ile Gln Asp Pro Val 820 825 830 Pro Gln PheArg Ile Asp Pro Asp Ser Gly Thr Met Tyr Thr Met Met 835 840 845 Glu LeuAsp Tyr Glu Asn Gln Val Ala Tyr Thr Leu Thr Ile Met Ala 850 855 860 GlnAsp Asn Gly Ile Pro Gln Lys Ser Asp Thr Thr Thr Leu Glu Ile 865 870 875880 Leu Ile Leu Asp Ala Asn Asp Asn Ala Pro Gln Phe Leu Trp Asp Phe 885890 895 Tyr Gln Gly Ser Ile Phe Glu Asp Ala Pro Pro Ser Thr Ser Ile Leu900 905 910 Gln Val Ser Ala Thr Asp Arg Asp Ser Gly Pro Asn Gly Arg LeuLeu 915 920 925 Tyr Thr Phe Gln Gly Gly Asp Asp Gly Asp Gly Asp Phe TyrIle Glu 930 935 940 Pro Thr Ser Gly Val Ile Arg Thr Gln Arg Arg Leu AspArg Glu Asn 945 950 955 960 Val Ala Val Tyr Asn Leu Trp Ala Leu Ala ValAsp Arg Gly Ser Pro 965 970 975 Thr Pro Leu Ser Ala Ser Val Glu Ile GlnVal Thr Ile Leu Asp Ile 980 985 990 Asn Asp Asn Ala Pro Met Phe Glu LysAsp Glu Leu Glu Leu Phe Val 995 1000 1005 Glu Glu Asn Asn Pro Val GlySer Val Val Ala Lys Ile Arg Ala Asn 1010 1015 1020 Asp Pro Asp Glu GlyPro Asn Ala Gln Ile Met Tyr Gln Ile Val Glu 1025 1030 1035 1040 Gly AspMet Arg His Phe Phe Gln Leu Asp Leu Leu Asn Gly Asp Leu 1045 1050 1055Arg Ala Met Val Glu Leu Asp Phe Glu Val Arg Arg Glu Tyr Val Leu 10601065 1070 Val Val Gln Ala Thr Ser Ala Pro Leu Val Ser Arg Ala Thr ValHis 1075 1080 1085 Ile Leu Leu Val Asp Gln Asn Asp Asn Pro Pro Val LeuPro Asp Phe 1090 1095 1100 Gln Ile Leu Phe Asn Asn Tyr Val Thr Asn LysSer Asn Ser Phe Pro 1105 1110 1115 1120 Thr Gly Val Ile Gly Cys Ile ProAla His Asp Pro Asp Val Ser Asp 1125 1130 1135 Ser Leu Asn Tyr Thr PheVal Gln Gly Asn Glu Leu Arg Leu Leu Leu 1140 1145 1150 Leu Asp Pro AlaThr Gly Glu Leu Gln Leu Ser Arg Asp Leu Asp Asn 1155 1160 1165 Asn ArgPro Leu Glu Ala Leu Met Glu Val Ser Val Ser Asp Gly Ile 1170 1175 1180His Ser Val Thr Ala Phe Cys Thr Leu Arg Val Thr Ile Ile Thr Asp 11851190 1195 1200 Asp Met Leu Thr Asn Ser Ile Thr Val Arg Leu Glu Asn MetSer Gln 1205 1210 1215 Glu Lys Phe Leu Ser Pro Leu Leu Ala Leu Phe ValGlu Gly Val Ala 1220 1225 1230 Ala Val Leu Ser Thr Thr Lys Asp Asp ValPhe Val Phe Asn Val Gln 1235 1240 1245 Asn Asp Thr Asp Val Ser Ser AsnIle Leu Asn Val Thr Phe Ser Ala 1250 1255 1260 Leu Leu Pro Gly Gly ValArg Gly Gln Phe Phe Pro Ser Glu Asp Leu 1265 1270 1275 1280 Gln Glu GlnIle Tyr Leu Asn Arg Thr Leu Leu Thr Thr Ile Ser Thr 1285 1290 1295 GlnArg Val Leu Pro Phe Asp Asp Asn Ile Cys Leu Arg Glu Pro Cys 1300 13051310 Glu Asn Tyr Met Lys Cys Val Ser Val Leu Arg Phe Asp Ser Ser Ala1315 1320 1325 Pro Phe Leu Ser Ser Thr Thr Val Leu Phe Arg Pro Ile HisPro Ile 1330 1335 1340 Asn Gly Leu Arg Cys Arg Cys Pro Pro Gly Phe ThrGly Asp Tyr Cys 1345 1350 1355 1360 Glu Thr Glu Ile Asp Leu Cys Tyr SerAsp Pro Cys Gly Ala Asn Gly 1365 1370 1375 Arg Cys Arg Ser Arg Glu GlyGly Tyr Thr Cys Glu Cys Phe Glu Asp 1380 1385 1390 Phe Thr Gly Glu HisCys Glu Val Asp Ala Arg Ser Gly Arg Cys Ala 1395 1400 1405 Asn Gly ValCys Lys Asn Gly Gly Thr Cys Val Asn Leu Leu Ile Gly 1410 1415 1420 GlyPhe His Cys Val Cys Pro Pro Gly Glu Tyr Glu Arg Pro Tyr Cys 1425 14301435 1440 Glu Val Thr Thr Arg Ser Phe Pro Pro Gln Ser Phe Val Thr PheArg 1445 1450 1455 Gly Leu Arg Gln Arg Phe His Phe Thr Ile Ser Leu ThrPhe Ala Thr 1460 1465 1470 Gln Glu Arg Asn Gly Leu Leu Leu Tyr Asn GlyArg Phe Asn Glu Lys 1475 1480 1485 His Asp Phe Ile Ala Leu Glu Ile ValAsp Glu Gln Val Gln Leu Thr 1490 1495 1500 Phe Ser Ala Gly Glu Thr ThrThr Thr Val Ala Pro Lys Val Pro Ser 1505 1510 1515 1520 Gly Val Ser AspGly Arg Trp His Ser Val Gln Val Gln Tyr Tyr Asn 1525 1530 1535 Lys ProAsn Ile Gly His Leu Gly Leu Pro His Gly Pro Ser Gly Glu 1540 1545 1550Lys Met Ala Val Val Thr Val Asp Asp Cys Asp Thr Thr Met Ala Val 15551560 1565 Arg Phe Gly Lys Asp Ile Gly Asn Tyr Ser Cys Ala Ala Gln GlyThr 1570 1575 1580 Gln Thr Gly Ser Lys Lys Ser Leu Asp Leu Thr Gly ProLeu Leu Leu 1585 1590 1595 1600 Gly Gly Val Pro Asn Leu Pro Glu Asp PhePro Val His Asn Arg Gln 1605 1610 1615 Phe Val Gly Cys Met Arg Asn LeuSer Val Asp Gly Lys Asn Val Asp 1620 1625 1630 Met Ala Gly Phe Ile AlaAsn Asn Gly Thr Arg Glu Gly Cys Ala Ala 1635 1640 1645 Arg Arg Asn PheCys Asp Gly Arg Arg Cys Gln Asn Gly Gly Thr Cys 1650 1655 1660 Val AsnArg Trp Asn Met Tyr Leu Cys Glu Cys Pro Leu Arg Phe Gly 1665 1670 16751680 Gly Lys Asn Cys Glu Gln Ala Met Pro His Pro Gln Leu Phe Ser Gly1685 1690 1695 Glu Ser Val Val Ser Trp Ser Asp Leu Asn Ile Ile Ile SerVal Pro 1700 1705 1710 Trp Tyr Leu Gly Leu Met Phe Arg Thr Arg Lys GluAsp Ser Val Leu 1715 1720 1725 Met Glu Ala Thr Ser Gly Gly Pro Thr SerPhe Arg Leu Gln Ile Leu 1730 1735 1740 Asn Asn Tyr Leu Gln Phe Glu ValSer His Gly Pro Ser Asp Val Glu 1745 1750 1755 1760 Ser Val Met Leu SerGly Leu Arg Val Thr Asp Gly Glu Trp His His 1765 1770 1775 Leu Leu IleGlu Leu Lys Asn Val Lys Glu Asp Ser Glu Met Lys His 1780 1785 1790 LeuVal Thr Met Thr Leu Asp Tyr Gly Met Asp Gln Asn Lys Ala Asp 1795 18001805 Ile Gly Gly Met Leu Pro Gly Leu Thr Val Arg Ser Val Val Val Gly1810 1815 1820 Gly Ala Ser Glu Asp Lys Val Ser Val Arg Arg Gly Phe ArgGly Cys 1825 1830 1835 1840 Met Gln Gly Val Arg Met Gly Gly Thr Pro ThrAsn Val Ala Thr Leu 1845 1850 1855 Asn Met Asn Asn Ala Leu Lys Val ArgVal Lys Asp Gly Cys Asp Val 1860 1865 1870 Asp Asp Pro Cys Thr Ser SerPro Cys Pro Pro Asn Ser Arg Cys His 1875 1880 1885 Asp Ala Trp Glu AspTyr Ser Cys Val Cys Asp Lys Gly Tyr Leu Gly 1890 1895 1900 Ile Asn CysVal Asp Ala Cys His Leu Asn Pro Cys Glu Asn Met Gly 1905 1910 1915 1920Ala Cys Val Arg Ser Pro Gly Ser Pro Gln Gly Tyr Val Cys Glu Cys 19251930 1935 Gly Pro Ser His Tyr Gly Pro Tyr Cys Glu Asn Lys Leu Asp LeuPro 1940 1945 1950 Cys Pro Arg Gly Trp Trp Gly Asn Pro Val Cys Gly ProCys His Cys 1955 1960 1965 Ala Val Ser Lys Gly Phe Asp Pro Asp Cys AsnLys Thr Asn Gly Gln 1970 1975 1980 Cys Gln Cys Lys Glu Asn Tyr Tyr LysLeu Leu Ala Gln Asp Thr Cys 1985 1990 1995 2000 Leu Pro Cys Asp Cys PhePro His Gly Ser His Ser Arg Thr Cys Asp 2005 2010 2015 Met Ala Thr GlyGln Cys Ala Cys Lys Pro Gly Val Ile Gly Arg Gln 2020 2025 2030 Cys AsnArg Cys Asp Asn Pro Phe Ala Glu Val Thr Thr Leu Gly Cys 2035 2040 2045Glu Val Ile Tyr Asn Gly Cys Pro Lys Ala Phe Glu Ala Gly Ile Trp 20502055 2060 Trp Pro Gln Thr Lys Phe Gly Gln Pro Ala Ala Val Pro Cys ProLys 2065 2070 2075 2080 Gly Ser Val Gly Asn Ala Val Arg His Cys Ser GlyGlu Lys Gly Trp 2085 2090 2095 Leu Pro Pro Glu Leu Phe Asn Cys Thr ThrIle Ser Phe Val Asp Leu 2100 2105 2110 Arg Ala Met Asn Glu Lys Leu SerArg Asn Glu Thr Gln Val Asp Gly 2115 2120 2125 Ala Arg Ala Leu Gln LeuVal Arg Ala Leu Arg Ser Ala Thr Gln His 2130 2135 2140 Thr Gly Thr LeuPhe Gly Asn Asp Val Arg Thr Ala Tyr Gln Leu Leu 2145 2150 2155 2160 GlyHis Val Leu Gln His Glu Ser Trp Gln Gln Gly Phe Asp Leu Ala 2165 21702175 Ala Thr Gln Asp Ala Asp Phe His Glu Asp Val Ile His Ser Gly Ser2180 2185 2190 Ala Leu Leu Ala Pro Ala Thr Arg Ala Ala Trp Glu Gln IleGln Arg 2195 2200 2205 Ser Glu Gly Gly Thr Ala Gln Leu Leu Arg Arg LeuGlu Gly Tyr Phe 2210 2215 2220 Ser Asn Val Ala Arg Asn Val Arg Arg ThrTyr Leu Arg Pro Phe Val 2225 2230 2235 2240 Ile Val Thr Ala Asn Met IleLeu Ala Val Asp Ile Phe Asp Lys Phe 2245 2250 2255 Asn Phe Thr Gly AlaArg Val Pro Arg Phe Asp Thr Ile His Glu Glu 2260 2265 2270 Phe Pro ArgGlu Leu Glu Ser Ser Val Ser Phe Pro Ala Asp Phe Phe 2275 2280 2285 ArgPro Pro Glu Glu Lys Glu Gly Pro Leu Leu Arg Pro Ala Gly Arg 2290 22952300 Arg Thr Thr Pro Gln Thr Thr Arg Pro Gly Pro Gly Thr Glu Arg Glu2305 2310 2315 2320 Ala Pro Ile Ser Arg Arg Arg Arg His Pro Asp Asp AlaGly Gln Phe 2325 2330 2335 Ala Val Ala Leu Val Ile Ile Tyr Arg Thr LeuGly Gln Leu Leu Pro 2340 2345 2350 Glu Arg Tyr Asp Pro Asp Arg Arg SerLeu Arg Leu Pro His Arg Pro 2355 2360 2365 Ile Ile Asn Thr Pro Met ValSer Thr Leu Val Tyr Ser Glu Gly Ala 2370 2375 2380 Pro Leu Pro Arg ProLeu Glu Arg Pro Val Leu Val Glu Phe Ala Leu 2385 2390 2395 2400 Leu GluVal Glu Glu Arg Thr Lys Pro Val Cys Val Phe Trp Asn His 2405 2410 2415Ser Leu Ala Val Gly Gly Thr Gly Gly Trp Ser Ala Arg Gly Cys Glu 24202425 2430 Leu Leu Ser Arg Asn Arg Thr His Val Ala Cys Gln Cys Ser HisThr 2435 2440 2445 Ala Ser Phe Ala Val Leu Met Asp Ile Ser Arg Arg GluAsn Gly Glu 2450 2455 2460 Val Leu Pro Leu Lys Ile Val Thr Tyr Ala AlaVal Ser Leu Ser Leu 2465 2470 2475 2480 Ala Ala Leu Leu Val Ala Phe ValLeu Leu Ser Leu Val Arg Met Leu 2485 2490 2495 Arg Ser Asn Leu His SerIle His Lys His Leu Ala Val Ala Leu Phe 2500 2505 2510 Leu Ser Gln LeuVal Phe Val Ile Gly Ile Asn Gln Thr Glu Asn Pro 2515 2520 2525 Phe LeuCys Thr Val Val Ala Ile Leu Leu His Tyr Ile Tyr Met Ser 2530 2535 2540Thr Phe Ala Trp Thr Leu Val Glu Ser Leu His Val Tyr Arg Met Leu 25452550 2555 2560 Thr Glu Val Arg Asn Ile Asp Thr Gly Pro Met Arg Phe TyrTyr Val 2565 2570 2575 Val Gly Trp Gly Ile Pro Ala Ile Val Thr Gly LeuAla Val Gly Leu 2580 2585 2590 Asp Pro Gln Gly Tyr Gly Asn Pro Asp PheCys Trp Leu Ser Leu Gln 2595 2600 2605 Asp Thr Leu Ile Trp Ser Phe AlaGly Pro Ile Gly Ala Val Ile Ile 2610 2615 2620 Ile Asn Thr Val Thr SerVal Leu Ser Ala Lys Val Ser Cys Gln Arg 2625 2630 2635 2640 Lys His HisTyr Tyr Gly Lys Lys Gly Ile Val Ser Leu Leu Arg Thr 2645 2650 2655 AlaPhe Leu Leu Leu Leu Leu Ile Ser Ala Thr Trp Leu Leu Gly Leu 2660 26652670 Leu Ala Val Asn Arg Asp Ala Leu Ser Phe His Tyr Leu Phe Ala Ile2675 2680 2685 Phe Ser Gly Leu Gln Gly Pro Phe Val Leu Leu Phe His CysVal Leu 2690 2695 2700 Asn Gln Glu Val Arg Lys His Leu Lys Gly Val LeuGly Gly Arg Lys 2705 2710 2715 2720 Leu His Leu Glu Asp Ser Ala Thr ThrArg Ala Thr Leu Leu Thr Arg 2725 2730 2735 Ser Leu Asn Cys Asn Thr ThrPhe Gly Asp Gly Pro Asp Met Leu Arg 2740 2745 2750 Thr Asp Leu Gly GluSer Thr Ala Ser Leu Asp Ser Ile Val Arg Asp 2755 2760 2765 Glu Gly IleGln Lys Leu Gly Val Ser Ser Gly Leu Val Arg Gly Ser 2770 2775 2780 HisGly Glu Pro Asp Ala Ser Leu Met Pro Arg Ser Cys Lys Asp Pro 2785 27902795 2800 Pro Gly His Asp Ser Asp Ser Asp Ser Glu Leu Ser Leu Asp GluGln 2805 2810 2815 Ser Ser Ser Tyr Ala Ser Ser His Ser Ser Asp Ser GluAsp Asp Gly 2820 2825 2830 Val Gly Ala Glu Glu Lys Trp Asp Pro Ala ArgGly Ala Val His Ser 2835 2840 2845 Thr Pro Lys Gly Asp Ala Val Ala AsnHis Val Pro Ala Gly Trp Pro 2850 2855 2860 Asp Gln Ser Leu Ala Glu SerAsp Ser Glu Asp Pro Ser Gly Lys Pro 2865 2870 2875 2880 Arg Leu Lys ValGlu Thr Lys Val Ser Val Glu Leu His Arg Glu Glu 2885 2890 2895 Gln GlySer His Arg Gly Glu Tyr Pro Pro Asp Gln Glu Ser Gly Gly 2900 2905 2910Ala Ala Arg Leu Ala Ser Ser Gln Pro Pro Glu Gln Arg Lys Gly Ile 29152920 2925 Leu Lys Asn Lys Val Thr Tyr Pro Pro Pro Leu Thr Leu Thr GluGln 2930 2935 2940 Thr Leu Lys Gly Arg Leu Arg Glu Lys Leu Ala Asp CysGlu Gln Ser 2945 2950 2955 2960 Pro Thr Ser Ser Arg Thr Ser Ser Leu GlySer Gly Gly Pro Asp Cys 2965 2970 2975 Ala Ile Thr Val Lys Ser Pro GlyArg Glu Pro Gly Arg Asp His Leu 2980 2985 2990 Asn Gly Val Ala Met AsnVal Arg Thr Gly Ser Ala Gln Ala Asp Gly 2995 3000 3005 Ser Asp Ser GluLys Pro 3010 3 2898 DNA Homo sapiens CDS (1)..(2898) 3 atg gag ttt gtgcgg gcg ctg tgg ctg ggc ctg gcg ctg gcg ctg ggg 48 Met Glu Phe Val ArgAla Leu Trp Leu Gly Leu Ala Leu Ala Leu Gly 1 5 10 15 ccg ggg tcc gcgggg ggc cac cct cag ccg tgc ggc gtc ctg gcg cgc 96 Pro Gly Ser Ala GlyGly His Pro Gln Pro Cys Gly Val Leu Ala Arg 20 25 30 ctc ggg ggc tcc gtgcgc ctg ggc gcc ctc ctg ccc cgc gcg cct ctc 144 Leu Gly Gly Ser Val ArgLeu Gly Ala Leu Leu Pro Arg Ala Pro Leu 35 40 45 gcc cgc gcc cgc gcc cgcgcc gcc ctg gcc cgg gcc gcc ctg gcg ccg 192 Ala Arg Ala Arg Ala Arg AlaAla Leu Ala Arg Ala Ala Leu Ala Pro 50 55 60 cgg ctg ccg cac aac ctg agcttg gag ctg gtg gtc gcc gcg ccc ccc 240 Arg Leu Pro His Asn Leu Ser LeuGlu Leu Val Val Ala Ala Pro Pro 65 70 75 80 gcc cgc gac ccc gcc tcg ctgacc cgc ggc ctg tgc cag gcg ctg gtg 288 Ala Arg Asp Pro Ala Ser Leu ThrArg Gly Leu Cys Gln Ala Leu Val 85 90 95 cct ccg ggc gtg gcg gcc ctg ctcgcc ttt ccc gag gct cgg ccc gag 336 Pro Pro Gly Val Ala Ala Leu Leu AlaPhe Pro Glu Ala Arg Pro Glu 100 105 110 ctg ctg cag ctg cac ttc ctg gcggcg gcc acc gag acc ccc gtg ctc 384 Leu Leu Gln Leu His Phe Leu Ala AlaAla Thr Glu Thr Pro Val Leu 115 120 125 agc ctg ctg cgg cgg gag gcg cgcgcg ccc ctc gga gcc ccg aac cca 432 Ser Leu Leu Arg Arg Glu Ala Arg AlaPro Leu Gly Ala Pro Asn Pro 130 135 140 ttc cac ctg cag ctg cac tgg gccagc ccc ctg gag acg ctg ctg gat 480 Phe His Leu Gln Leu His Trp Ala SerPro Leu Glu Thr Leu Leu Asp 145 150 155 160 gtg ctg gtg gcg gtg ctg caggcg cac gcc tgg gaa gac gtc ggc ctg 528 Val Leu Val Ala Val Leu Gln AlaHis Ala Trp Glu Asp Val Gly Leu 165 170 175 gcc ctg tgc cgc act cag gacccc ggc ggc ctg gtg gcc ctc tgg aca 576 Ala Leu Cys Arg Thr Gln Asp ProGly Gly Leu Val Ala Leu Trp Thr 180 185 190 agc cgg gct ggc cgg ccc ccacag ctg gtc ctg gac cta agc cgg cgg 624 Ser Arg Ala Gly Arg Pro Pro GlnLeu Val Leu Asp Leu Ser Arg Arg 195 200 205 gac acg gga gat gca gga ctgcgg gca cgc ctg gcc ccg atg gcg gcg 672 Asp Thr Gly Asp Ala Gly Leu ArgAla Arg Leu Ala Pro Met Ala Ala 210 215 220 cca gtg ggg ggt gaa gca ccggta ccc gcg gcg gtc ctc ctc ggc tgt 720 Pro Val Gly Gly Glu Ala Pro ValPro Ala Ala Val Leu Leu Gly Cys 225 230 235 240 gac atc gcc cgt gcc cgtcgg gtg ctg gag gcc gta cct ccc ggc ccc 768 Asp Ile Ala Arg Ala Arg ArgVal Leu Glu Ala Val Pro Pro Gly Pro 245 250 255 cac tgg ctg ttg ggg acacca ctg ccg ccc aag gcc ctg ccc acc gcg 816 His Trp Leu Leu Gly Thr ProLeu Pro Pro Lys Ala Leu Pro Thr Ala 260 265 270 ggg ctg cca cca ggg ctgctg gcg ctg ggc gag gtg gca cga ccc ccg 864 Gly Leu Pro Pro Gly Leu LeuAla Leu Gly Glu Val Ala Arg Pro Pro 275 280 285 ctg gag gcc gcc atc catgac att gtg caa ctg gtg gcc cgg gcg ctg 912 Leu Glu Ala Ala Ile His AspIle Val Gln Leu Val Ala Arg Ala Leu 290 295 300 ggc agt gcg gcc cag gtgcag ccg aag cga gcc ctc ctc ccc gcc ccg 960 Gly Ser Ala Ala Gln Val GlnPro Lys Arg Ala Leu Leu Pro Ala Pro 305 310 315 320 gtc aac tgc ggg gacctg cag ccg gcc ggg ccc gag tcc ccg ggg cgc 1008 Val Asn Cys Gly Asp LeuGln Pro Ala Gly Pro Glu Ser Pro Gly Arg 325 330 335 ttc ttg gca cgg ttcctg gcc aac acg tcc ttc cag ggc cgc acg ggc 1056 Phe Leu Ala Arg Phe LeuAla Asn Thr Ser Phe Gln Gly Arg Thr Gly 340 345 350 ccc gtg tgg gtg acaggc agc tcc cca gac gaa gac ggg cag tgc cca 1104 Pro Val Trp Val Thr GlySer Ser Pro Asp Glu Asp Gly Gln Cys Pro 355 360 365 gcg ggg cag ctg tgcctg gac cct ggc acc aac gac tcg gcc acc ctg 1152 Ala Gly Gln Leu Cys LeuAsp Pro Gly Thr Asn Asp Ser Ala Thr Leu 370 375 380 gac gca ctg ttc gccgcg ctg gcc aac ggc tca gcg ccc cgt gcc ctg 1200 Asp Ala Leu Phe Ala AlaLeu Ala Asn Gly Ser Ala Pro Arg Ala Leu 385 390 395 400 cgc aag tgc tgctac ggc tac tgc att gac ctg ctg gag cgg ctg gcg 1248 Arg Lys Cys Cys TyrGly Tyr Cys Ile Asp Leu Leu Glu Arg Leu Ala 405 410 415 gag gac acg cccttc gac ttc gag ctg tac ctc gtg ggt gac ggc aag 1296 Glu Asp Thr Pro PheAsp Phe Glu Leu Tyr Leu Val Gly Asp Gly Lys 420 425 430 tac ggc gcc ctgcgg gac ggc cgc tgg acc ggc ctg gtc ggg gac ctg 1344 Tyr Gly Ala Leu ArgAsp Gly Arg Trp Thr Gly Leu Val Gly Asp Leu 435 440 445 ctg gcc ggc cgggcc cac atg gcg gtc acc agc ttc agt atc aac tcc 1392 Leu Ala Gly Arg AlaHis Met Ala Val Thr Ser Phe Ser Ile Asn Ser 450 455 460 gcc cgc tca caggtg gtg gac ttc acc agc ccc ttc ttc tcc acc agc 1440 Ala Arg Ser Gln ValVal Asp Phe Thr Ser Pro Phe Phe Ser Thr Ser 465 470 475 480 ctg ggc atcatg gtg cgg gca cgg gac acg gcc tca ccc atc ggt gcc 1488 Leu Gly Ile MetVal Arg Ala Arg Asp Thr Ala Ser Pro Ile Gly Ala 485 490 495 ttt atg tggccc ctg cac tgg tcc acg tgg ctg ggc gtc ttt gcg gcc 1536 Phe Met Trp ProLeu His Trp Ser Thr Trp Leu Gly Val Phe Ala Ala 500 505 510 ctg cac ctcacc gcg ctc ttc ctc acc gtg tac gag tgg cgt agc ccc 1584 Leu His Leu ThrAla Leu Phe Leu Thr Val Tyr Glu Trp Arg Ser Pro 515 520 525 tac ggc ctcacg cca cgt ggc cgc aac cgc agc acc gtc ttc tcc tac 1632 Tyr Gly Leu ThrPro Arg Gly Arg Asn Arg Ser Thr Val Phe Ser Tyr 530 535 540 tcc tca gccctc aac ctg tgc tac gcc atc ctc ttc aga cgc acc gtg 1680 Ser Ser Ala LeuAsn Leu Cys Tyr Ala Ile Leu Phe Arg Arg Thr Val 545 550 555 560 tcc agcaag acg ccc aag tgc ccc acg ggc cgc ctg ctc atg aac ctc 1728 Ser Ser LysThr Pro Lys Cys Pro Thr Gly Arg Leu Leu Met Asn Leu 565 570 575 tgg gccatc ttc tgc ctg ctg gtg ctg tcc agc tac acg gcc aac ctg 1776 Trp Ala IlePhe Cys Leu Leu Val Leu Ser Ser Tyr Thr Ala Asn Leu 580 585 590 gct gccgtc atg gtc ggg gac aag acc ttc gag gag ctg tcg ggg atc 1824 Ala Ala ValMet Val Gly Asp Lys Thr Phe Glu Glu Leu Ser Gly Ile 595 600 605 cac gacccc aag ggc ttc cgc ttc ggc acc gtg tgg gag agc agc gcc 1872 His Asp ProLys Gly Phe Arg Phe Gly Thr Val Trp Glu Ser Ser Ala 610 615 620 gag gcgtac atc aag aag agc ttc ccc gac atg cac gca cac atg cgg 1920 Glu Ala TyrIle Lys Lys Ser Phe Pro Asp Met His Ala His Met Arg 625 630 635 640 cgccac agc gcg ccc acc acg ccc cgc ggc gtc gcc atg ctc acg agc 1968 Arg HisSer Ala Pro Thr Thr Pro Arg Gly Val Ala Met Leu Thr Ser 645 650 655 gacccc ccc aag ctc aac gcc ttc atc atg gac aag tcg ctc ctg gac 2016 Asp ProPro Lys Leu Asn Ala Phe Ile Met Asp Lys Ser Leu Leu Asp 660 665 670 tacgag gtc tcc atc gac gcc gac tgc aaa ctg ctg acc gtg gga aag 2064 Tyr GluVal Ser Ile Asp Ala Asp Cys Lys Leu Leu Thr Val Gly Lys 675 680 685 cccttc gcc att gag ggc tat ggg atc gga ctg ccc cag aac tcg ccg 2112 Pro PheAla Ile Glu Gly Tyr Gly Ile Gly Leu Pro Gln Asn Ser Pro 690 695 700 ctcacc tcc aac ctg tcc gag ttc atc agc cgc tac aag tcc tcc ggc 2160 Leu ThrSer Asn Leu Ser Glu Phe Ile Ser Arg Tyr Lys Ser Ser Gly 705 710 715 720ttc atc gac ctg ctc cac gac aag tgg tac aag atg gtg cct tgc ggc 2208 PheIle Asp Leu Leu His Asp Lys Trp Tyr Lys Met Val Pro Cys Gly 725 730 735aag cgg gtc ttt gcg gtt aca gag acc ctg cag atg agc atc tac cac 2256 LysArg Val Phe Ala Val Thr Glu Thr Leu Gln Met Ser Ile Tyr His 740 745 750ttc gcg ggc ctc ttc gtg ttg ctg tgc ctg ggc ctg ggc agc gct ctg 2304 PheAla Gly Leu Phe Val Leu Leu Cys Leu Gly Leu Gly Ser Ala Leu 755 760 765ctc agc tcg ctg ggc gag cac gcc ttc ttc cgc ctg gcg ctg ccg cgc 2352 LeuSer Ser Leu Gly Glu His Ala Phe Phe Arg Leu Ala Leu Pro Arg 770 775 780atc cgc aag ggg agc agg ctg cag tac tgg ctg cac acc agc cag aaa 2400 IleArg Lys Gly Ser Arg Leu Gln Tyr Trp Leu His Thr Ser Gln Lys 785 790 795800 atc cac cgc gcc ctc aac acg gag cca cca gag ggg tcg aag gag gag 2448Ile His Arg Ala Leu Asn Thr Glu Pro Pro Glu Gly Ser Lys Glu Glu 805 810815 acg gca gag gcg gag ccc agc ggc ccc gag gtg gag cag cag cag cag 2496Thr Ala Glu Ala Glu Pro Ser Gly Pro Glu Val Glu Gln Gln Gln Gln 820 825830 cag cag gac cag cca acg gct ccg gag ggc tgg aaa cgg gcg cgc cgg 2544Gln Gln Asp Gln Pro Thr Ala Pro Glu Gly Trp Lys Arg Ala Arg Arg 835 840845 gcc gtg gac aag gag cgc cgc gtg cgc ttc ctg ctg gag ccc gcc gtg 2592Ala Val Asp Lys Glu Arg Arg Val Arg Phe Leu Leu Glu Pro Ala Val 850 855860 gtt gtg gca ccc gaa gcg gac gcg gag gcg gag gct gcg ccg cga gag 2640Val Val Ala Pro Glu Ala Asp Ala Glu Ala Glu Ala Ala Pro Arg Glu 865 870875 880 ggc ccc gtc tgg ctg tgc tcc tac ggc cgc ccg ccc gcc gca agg ccc2688 Gly Pro Val Trp Leu Cys Ser Tyr Gly Arg Pro Pro Ala Ala Arg Pro 885890 895 acg ggg gcc ccc cag ccc ggg gag ctg cag gag ctg gag cgc cgc atc2736 Thr Gly Ala Pro Gln Pro Gly Glu Leu Gln Glu Leu Glu Arg Arg Ile 900905 910 gaa gtc gcg cgt gag cgg ctc cgc cag gcc ctg gtg cgg cgc ggc cag2784 Glu Val Ala Arg Glu Arg Leu Arg Gln Ala Leu Val Arg Arg Gly Gln 915920 925 ctc ctg gca cag ctc ggg gac agc gca cgt cac cgg cct cgg cgc ttg2832 Leu Leu Ala Gln Leu Gly Asp Ser Ala Arg His Arg Pro Arg Arg Leu 930935 940 ctt cag gcc aga gcg gcc ccc gcg gag gcc cca cca cac tct ggc cga2880 Leu Gln Ala Arg Ala Ala Pro Ala Glu Ala Pro Pro His Ser Gly Arg 945950 955 960 ccg ggg agc cag gaa tga 2898 Pro Gly Ser Gln Glu 965 4 965PRT Homo sapiens 4 Met Glu Phe Val Arg Ala Leu Trp Leu Gly Leu Ala LeuAla Leu Gly 1 5 10 15 Pro Gly Ser Ala Gly Gly His Pro Gln Pro Cys GlyVal Leu Ala Arg 20 25 30 Leu Gly Gly Ser Val Arg Leu Gly Ala Leu Leu ProArg Ala Pro Leu 35 40 45 Ala Arg Ala Arg Ala Arg Ala Ala Leu Ala Arg AlaAla Leu Ala Pro 50 55 60 Arg Leu Pro His Asn Leu Ser Leu Glu Leu Val ValAla Ala Pro Pro 65 70 75 80 Ala Arg Asp Pro Ala Ser Leu Thr Arg Gly LeuCys Gln Ala Leu Val 85 90 95 Pro Pro Gly Val Ala Ala Leu Leu Ala Phe ProGlu Ala Arg Pro Glu 100 105 110 Leu Leu Gln Leu His Phe Leu Ala Ala AlaThr Glu Thr Pro Val Leu 115 120 125 Ser Leu Leu Arg Arg Glu Ala Arg AlaPro Leu Gly Ala Pro Asn Pro 130 135 140 Phe His Leu Gln Leu His Trp AlaSer Pro Leu Glu Thr Leu Leu Asp 145 150 155 160 Val Leu Val Ala Val LeuGln Ala His Ala Trp Glu Asp Val Gly Leu 165 170 175 Ala Leu Cys Arg ThrGln Asp Pro Gly Gly Leu Val Ala Leu Trp Thr 180 185 190 Ser Arg Ala GlyArg Pro Pro Gln Leu Val Leu Asp Leu Ser Arg Arg 195 200 205 Asp Thr GlyAsp Ala Gly Leu Arg Ala Arg Leu Ala Pro Met Ala Ala 210 215 220 Pro ValGly Gly Glu Ala Pro Val Pro Ala Ala Val Leu Leu Gly Cys 225 230 235 240Asp Ile Ala Arg Ala Arg Arg Val Leu Glu Ala Val Pro Pro Gly Pro 245 250255 His Trp Leu Leu Gly Thr Pro Leu Pro Pro Lys Ala Leu Pro Thr Ala 260265 270 Gly Leu Pro Pro Gly Leu Leu Ala Leu Gly Glu Val Ala Arg Pro Pro275 280 285 Leu Glu Ala Ala Ile His Asp Ile Val Gln Leu Val Ala Arg AlaLeu 290 295 300 Gly Ser Ala Ala Gln Val Gln Pro Lys Arg Ala Leu Leu ProAla Pro 305 310 315 320 Val Asn Cys Gly Asp Leu Gln Pro Ala Gly Pro GluSer Pro Gly Arg 325 330 335 Phe Leu Ala Arg Phe Leu Ala Asn Thr Ser PheGln Gly Arg Thr Gly 340 345 350 Pro Val Trp Val Thr Gly Ser Ser Pro AspGlu Asp Gly Gln Cys Pro 355 360 365 Ala Gly Gln Leu Cys Leu Asp Pro GlyThr Asn Asp Ser Ala Thr Leu 370 375 380 Asp Ala Leu Phe Ala Ala Leu AlaAsn Gly Ser Ala Pro Arg Ala Leu 385 390 395 400 Arg Lys Cys Cys Tyr GlyTyr Cys Ile Asp Leu Leu Glu Arg Leu Ala 405 410 415 Glu Asp Thr Pro PheAsp Phe Glu Leu Tyr Leu Val Gly Asp Gly Lys 420 425 430 Tyr Gly Ala LeuArg Asp Gly Arg Trp Thr Gly Leu Val Gly Asp Leu 435 440 445 Leu Ala GlyArg Ala His Met Ala Val Thr Ser Phe Ser Ile Asn Ser 450 455 460 Ala ArgSer Gln Val Val Asp Phe Thr Ser Pro Phe Phe Ser Thr Ser 465 470 475 480Leu Gly Ile Met Val Arg Ala Arg Asp Thr Ala Ser Pro Ile Gly Ala 485 490495 Phe Met Trp Pro Leu His Trp Ser Thr Trp Leu Gly Val Phe Ala Ala 500505 510 Leu His Leu Thr Ala Leu Phe Leu Thr Val Tyr Glu Trp Arg Ser Pro515 520 525 Tyr Gly Leu Thr Pro Arg Gly Arg Asn Arg Ser Thr Val Phe SerTyr 530 535 540 Ser Ser Ala Leu Asn Leu Cys Tyr Ala Ile Leu Phe Arg ArgThr Val 545 550 555 560 Ser Ser Lys Thr Pro Lys Cys Pro Thr Gly Arg LeuLeu Met Asn Leu 565 570 575 Trp Ala Ile Phe Cys Leu Leu Val Leu Ser SerTyr Thr Ala Asn Leu 580 585 590 Ala Ala Val Met Val Gly Asp Lys Thr PheGlu Glu Leu Ser Gly Ile 595 600 605 His Asp Pro Lys Gly Phe Arg Phe GlyThr Val Trp Glu Ser Ser Ala 610 615 620 Glu Ala Tyr Ile Lys Lys Ser PhePro Asp Met His Ala His Met Arg 625 630 635 640 Arg His Ser Ala Pro ThrThr Pro Arg Gly Val Ala Met Leu Thr Ser 645 650 655 Asp Pro Pro Lys LeuAsn Ala Phe Ile Met Asp Lys Ser Leu Leu Asp 660 665 670 Tyr Glu Val SerIle Asp Ala Asp Cys Lys Leu Leu Thr Val Gly Lys 675 680 685 Pro Phe AlaIle Glu Gly Tyr Gly Ile Gly Leu Pro Gln Asn Ser Pro 690 695 700 Leu ThrSer Asn Leu Ser Glu Phe Ile Ser Arg Tyr Lys Ser Ser Gly 705 710 715 720Phe Ile Asp Leu Leu His Asp Lys Trp Tyr Lys Met Val Pro Cys Gly 725 730735 Lys Arg Val Phe Ala Val Thr Glu Thr Leu Gln Met Ser Ile Tyr His 740745 750 Phe Ala Gly Leu Phe Val Leu Leu Cys Leu Gly Leu Gly Ser Ala Leu755 760 765 Leu Ser Ser Leu Gly Glu His Ala Phe Phe Arg Leu Ala Leu ProArg 770 775 780 Ile Arg Lys Gly Ser Arg Leu Gln Tyr Trp Leu His Thr SerGln Lys 785 790 795 800 Ile His Arg Ala Leu Asn Thr Glu Pro Pro Glu GlySer Lys Glu Glu 805 810 815 Thr Ala Glu Ala Glu Pro Ser Gly Pro Glu ValGlu Gln Gln Gln Gln 820 825 830 Gln Gln Asp Gln Pro Thr Ala Pro Glu GlyTrp Lys Arg Ala Arg Arg 835 840 845 Ala Val Asp Lys Glu Arg Arg Val ArgPhe Leu Leu Glu Pro Ala Val 850 855 860 Val Val Ala Pro Glu Ala Asp AlaGlu Ala Glu Ala Ala Pro Arg Glu 865 870 875 880 Gly Pro Val Trp Leu CysSer Tyr Gly Arg Pro Pro Ala Ala Arg Pro 885 890 895 Thr Gly Ala Pro GlnPro Gly Glu Leu Gln Glu Leu Glu Arg Arg Ile 900 905 910 Glu Val Ala ArgGlu Arg Leu Arg Gln Ala Leu Val Arg Arg Gly Gln 915 920 925 Leu Leu AlaGln Leu Gly Asp Ser Ala Arg His Arg Pro Arg Arg Leu 930 935 940 Leu GlnAla Arg Ala Ala Pro Ala Glu Ala Pro Pro His Ser Gly Arg 945 950 955 960Pro Gly Ser Gln Glu 965 5 2916 DNA Homo sapiens CDS (1)..(2916) 5 atggag ttt gtg cgg gcg ctg tgg ctg ggc ctg gcg ctg gcg ctg ggg 48 Met GluPhe Val Arg Ala Leu Trp Leu Gly Leu Ala Leu Ala Leu Gly 1 5 10 15 ccgggg tcc gcg ggg ggc cac cct cag ccg tgc ggc gtc ctg gcg cgc 96 Pro GlySer Ala Gly Gly His Pro Gln Pro Cys Gly Val Leu Ala Arg 20 25 30 ctc gggggc tcc gtg cgc ctg ggc gcc ctc ctg ccc cgc gcg cct ctc 144 Leu Gly GlySer Val Arg Leu Gly Ala Leu Leu Pro Arg Ala Pro Leu 35 40 45 gcc cgc gcccgc gcc cgc gcc gcc ctg gcc cgg gcc gcc ctg gcg ccg 192 Ala Arg Ala ArgAla Arg Ala Ala Leu Ala Arg Ala Ala Leu Ala Pro 50 55 60 cgg ctg ccg cacaac ctg agc ttg gag ctg gtg gtc gcc gcg ccc ccc 240 Arg Leu Pro His AsnLeu Ser Leu Glu Leu Val Val Ala Ala Pro Pro 65 70 75 80 gcc cgc gac cccgcc tcg ctg acc cgc ggc ctg tgc cag gcg ctg gtg 288 Ala Arg Asp Pro AlaSer Leu Thr Arg Gly Leu Cys Gln Ala Leu Val 85 90 95 cct ccg ggc gtg gcggcc ctg ctc gcc ttt ccc gag gct cgg ccc gag 336 Pro Pro Gly Val Ala AlaLeu Leu Ala Phe Pro Glu Ala Arg Pro Glu 100 105 110 ctg ctg cag ctg cacttc ctg gcg gcg gcc acc gag acc ccc gtg ctc 384 Leu Leu Gln Leu His PheLeu Ala Ala Ala Thr Glu Thr Pro Val Leu 115 120 125 agc ctg ctg cgg cgggag gcg cgc gcg ccc ctc gga gcc ccg aac cca 432 Ser Leu Leu Arg Arg GluAla Arg Ala Pro Leu Gly Ala Pro Asn Pro 130 135 140 ttc cac ctg cag ctgcac tgg gcc agc ccc ctg gag acg ctg ctg gat 480 Phe His Leu Gln Leu HisTrp Ala Ser Pro Leu Glu Thr Leu Leu Asp 145 150 155 160 gtg ctg gtg gcggtg ctg cag gcg cac gcc tgg gaa gac gtc ggc ctg 528 Val Leu Val Ala ValLeu Gln Ala His Ala Trp Glu Asp Val Gly Leu 165 170 175 gcc ctg tgc cgcact cag gac ccc ggc ggc ctg gtg gcc ctc tgg aca 576 Ala Leu Cys Arg ThrGln Asp Pro Gly Gly Leu Val Ala Leu Trp Thr 180 185 190 agc cgg gct ggccgg ccc cca cag ctg gtc ctg gac cta agc cgg cgg 624 Ser Arg Ala Gly ArgPro Pro Gln Leu Val Leu Asp Leu Ser Arg Arg 195 200 205 gac acg gga gatgca gga ctg cgg gca cgc ctg gcc ccg atg gcg gcg 672 Asp Thr Gly Asp AlaGly Leu Arg Ala Arg Leu Ala Pro Met Ala Ala 210 215 220 cca gtg ggg ggtgaa gca ccg gta ccc gcg gcg gtc ctc ctc ggc tgt 720 Pro Val Gly Gly GluAla Pro Val Pro Ala Ala Val Leu Leu Gly Cys 225 230 235 240 gac atc gcccgt gcc cgt cgg gtg ctg gag gcc gta cct ccc ggc ccc 768 Asp Ile Ala ArgAla Arg Arg Val Leu Glu Ala Val Pro Pro Gly Pro 245 250 255 cac tgg ctgttg ggg aca cca ctg ccg ccc aag gcc ctg ccc acc gcg 816 His Trp Leu LeuGly Thr Pro Leu Pro Pro Lys Ala Leu Pro Thr Ala 260 265 270 ggg ctg ccacca ggg ctg ctg gcg ctg ggc gag gtg gca cga ccc ccg 864 Gly Leu Pro ProGly Leu Leu Ala Leu Gly Glu Val Ala Arg Pro Pro 275 280 285 ctg gag gccgcc atc cat gac att gtg caa ctg gtg gcc cgg gcg ctg 912 Leu Glu Ala AlaIle His Asp Ile Val Gln Leu Val Ala Arg Ala Leu 290 295 300 ggc agt gcggcc cag gtg cag ccg aag cga gcc ctc ctc ccc gcc ccg 960 Gly Ser Ala AlaGln Val Gln Pro Lys Arg Ala Leu Leu Pro Ala Pro 305 310 315 320 gtc aactgc ggg gac ctg cag ccg gcc ggg ccc gag tcc ccg ggg cgc 1008 Val Asn CysGly Asp Leu Gln Pro Ala Gly Pro Glu Ser Pro Gly Arg 325 330 335 ttc ttggca cgg ttc ctg gcc aac acg tcc ttc cag ggc cgc acg ggc 1056 Phe Leu AlaArg Phe Leu Ala Asn Thr Ser Phe Gln Gly Arg Thr Gly 340 345 350 ccc gtgtgg gtg aca ggc agc tcc cca gac gaa gac ggg cag tgc cca 1104 Pro Val TrpVal Thr Gly Ser Ser Pro Asp Glu Asp Gly Gln Cys Pro 355 360 365 gcg gggcag ctg tgc ctg gac cct ggc acc aac gac tcg gcc acc ctg 1152 Ala Gly GlnLeu Cys Leu Asp Pro Gly Thr Asn Asp Ser Ala Thr Leu 370 375 380 gac gcactg ttc gcc gcg ctg gcc aac ggc tca gcg ccc cgt gcc ctg 1200 Asp Ala LeuPhe Ala Ala Leu Ala Asn Gly Ser Ala Pro Arg Ala Leu 385 390 395 400 cgcaag tgc tgc tac ggc tac tgc att gac ctg ctg gag cgg ctg gcg 1248 Arg LysCys Cys Tyr Gly Tyr Cys Ile Asp Leu Leu Glu Arg Leu Ala 405 410 415 gaggac acg ccc ttc gac ttc gag ctg tac ctc gtg ggt gac ggc aag 1296 Glu AspThr Pro Phe Asp Phe Glu Leu Tyr Leu Val Gly Asp Gly Lys 420 425 430 tacggc gcc ctg cgg gac ggc cgc tgg acc ggc ctg gtc ggg gac ctg 1344 Tyr GlyAla Leu Arg Asp Gly Arg Trp Thr Gly Leu Val Gly Asp Leu 435 440 445 ctggcc ggc cgg gcc cac atg gcg gtc acc agc ttc agt atc aac tcc 1392 Leu AlaGly Arg Ala His Met Ala Val Thr Ser Phe Ser Ile Asn Ser 450 455 460 gcccgc tca cag gtg gtg gac ttc acc agc ccc ttc ttc tcc acc agc 1440 Ala ArgSer Gln Val Val Asp Phe Thr Ser Pro Phe Phe Ser Thr Ser 465 470 475 480ctg ggc atc atg gtg cgg gca cgg gac acg gcc tca ccc atc ggt gcc 1488 LeuGly Ile Met Val Arg Ala Arg Asp Thr Ala Ser Pro Ile Gly Ala 485 490 495ttt atg tgg ccc ctg cac tgg tcc acg tgg ctg ggc gtc ttt gcg gcc 1536 PheMet Trp Pro Leu His Trp Ser Thr Trp Leu Gly Val Phe Ala Ala 500 505 510ctg cac ctc acc gcg ctc ttc ctc acc gtg tac gag tgg cgt agc ccc 1584 LeuHis Leu Thr Ala Leu Phe Leu Thr Val Tyr Glu Trp Arg Ser Pro 515 520 525tac ggc ctc acg cca cgt ggc cgc aac cgc agc acc gtc ttc tcc tac 1632 TyrGly Leu Thr Pro Arg Gly Arg Asn Arg Ser Thr Val Phe Ser Tyr 530 535 540tcc tca gcc ctc aac ctg tgc tac gcc atc ctc ttc aga cgc acc gtg 1680 SerSer Ala Leu Asn Leu Cys Tyr Ala Ile Leu Phe Arg Arg Thr Val 545 550 555560 tcc agc aag acg ccc aag tgc ccc acg ggc cgc ctg ctc atg aac ctc 1728Ser Ser Lys Thr Pro Lys Cys Pro Thr Gly Arg Leu Leu Met Asn Leu 565 570575 tgg gcc atc ttc tgc ctg ctg gtg ctg tcc agc tac acg gcc aac ctg 1776Trp Ala Ile Phe Cys Leu Leu Val Leu Ser Ser Tyr Thr Ala Asn Leu 580 585590 gct gcc gtc atg gtc ggg gac aag acc ttc gag gag ctg tcg ggg atc 1824Ala Ala Val Met Val Gly Asp Lys Thr Phe Glu Glu Leu Ser Gly Ile 595 600605 cac gac ccc aag ctg cac cac ccg gcg cag ggc ttc cgc ttc ggc acc 1872His Asp Pro Lys Leu His His Pro Ala Gln Gly Phe Arg Phe Gly Thr 610 615620 gtg tgg gag agc agc gcc gag gcg tac atc aag aag agc ttc ccc gac 1920Val Trp Glu Ser Ser Ala Glu Ala Tyr Ile Lys Lys Ser Phe Pro Asp 625 630635 640 atg cac gca cac atg cgg cgc cac agc gcg ccc acc acg ccc cgc ggc1968 Met His Ala His Met Arg Arg His Ser Ala Pro Thr Thr Pro Arg Gly 645650 655 gtc gcc atg ctc acg agc gac ccc ccc aag ctc aac gcc ttc atc atg2016 Val Ala Met Leu Thr Ser Asp Pro Pro Lys Leu Asn Ala Phe Ile Met 660665 670 gac aag tcg ctc ctg gac tac gag gtc tcc atc gac gcc gac tgc aaa2064 Asp Lys Ser Leu Leu Asp Tyr Glu Val Ser Ile Asp Ala Asp Cys Lys 675680 685 ctg ctg acc gtg gga aag ccc ttc gcc att gag ggc tat ggg atc gga2112 Leu Leu Thr Val Gly Lys Pro Phe Ala Ile Glu Gly Tyr Gly Ile Gly 690695 700 ctg ccc cag aac tcg ccg ctc acc tcc aac ctg tcc gag ttc atc agc2160 Leu Pro Gln Asn Ser Pro Leu Thr Ser Asn Leu Ser Glu Phe Ile Ser 705710 715 720 cgc tac aag tcc tcc ggc ttc atc gac ctg ctc cac gac aag tggtac 2208 Arg Tyr Lys Ser Ser Gly Phe Ile Asp Leu Leu His Asp Lys Trp Tyr725 730 735 aag atg gtg cct tgc ggc aag cgg gtc ttt gcg gtt aca gag accctg 2256 Lys Met Val Pro Cys Gly Lys Arg Val Phe Ala Val Thr Glu Thr Leu740 745 750 cag atg agc atc tac cac ttc gcg ggc ctc ttc gtg ttg ctg tgcctg 2304 Gln Met Ser Ile Tyr His Phe Ala Gly Leu Phe Val Leu Leu Cys Leu755 760 765 ggc ctg ggc agc gct ctg ctc agc tcg ctg ggc gag cac gcc ttcttc 2352 Gly Leu Gly Ser Ala Leu Leu Ser Ser Leu Gly Glu His Ala Phe Phe770 775 780 cgc ctg gcg ctg ccg cgc atc cgc aag ggg agc agg ctg cag tactgg 2400 Arg Leu Ala Leu Pro Arg Ile Arg Lys Gly Ser Arg Leu Gln Tyr Trp785 790 795 800 ctg cac acc agc cag aaa atc cac cgc gcc ctc aac acg gagcca cca 2448 Leu His Thr Ser Gln Lys Ile His Arg Ala Leu Asn Thr Glu ProPro 805 810 815 gag ggg tcg aag gag gag acg gca gag gcg gag ccc agc ggcccc gag 2496 Glu Gly Ser Lys Glu Glu Thr Ala Glu Ala Glu Pro Ser Gly ProGlu 820 825 830 gtg gag cag cag cag cag cag cag gac cag cca acg gct ccggag ggc 2544 Val Glu Gln Gln Gln Gln Gln Gln Asp Gln Pro Thr Ala Pro GluGly 835 840 845 tgg aaa cgg gcg cgc cgg gcc gtg gac aag gag cgc cgc gtgcgc ttc 2592 Trp Lys Arg Ala Arg Arg Ala Val Asp Lys Glu Arg Arg Val ArgPhe 850 855 860 ctg ctg gag ccc gcc gtg gtt gtg gca ccc gaa gcg gac gcggag gcg 2640 Leu Leu Glu Pro Ala Val Val Val Ala Pro Glu Ala Asp Ala GluAla 865 870 875 880 gag gct gcg ccg cga gag ggc ccc gtc tgg ctg tgc tcctac ggc cgc 2688 Glu Ala Ala Pro Arg Glu Gly Pro Val Trp Leu Cys Ser TyrGly Arg 885 890 895 ccg ccc gcc gca agg ccc acg ggg gcc ccc cag ccc ggggag ctg cag 2736 Pro Pro Ala Ala Arg Pro Thr Gly Ala Pro Gln Pro Gly GluLeu Gln 900 905 910 gag ctg gag cgc cgc atc gaa gtc gcg cgt gag cgg ctccgc cag gcc 2784 Glu Leu Glu Arg Arg Ile Glu Val Ala Arg Glu Arg Leu ArgGln Ala 915 920 925 ctg gtg cgg cgc ggc cag ctc ctg gca cag ctc ggg gacagc gca cgt 2832 Leu Val Arg Arg Gly Gln Leu Leu Ala Gln Leu Gly Asp SerAla Arg 930 935 940 cac cgg cct cgg cgc ttg ctt cag gcc aga gcg gcc cccgcg gag gcc 2880 His Arg Pro Arg Arg Leu Leu Gln Ala Arg Ala Ala Pro AlaGlu Ala 945 950 955 960 cca cca cac tct ggc cga ccg ggg agc cag gaa tga2916 Pro Pro His Ser Gly Arg Pro Gly Ser Gln Glu 965 970 6 971 PRT Homosapiens 6 Met Glu Phe Val Arg Ala Leu Trp Leu Gly Leu Ala Leu Ala LeuGly 1 5 10 15 Pro Gly Ser Ala Gly Gly His Pro Gln Pro Cys Gly Val LeuAla Arg 20 25 30 Leu Gly Gly Ser Val Arg Leu Gly Ala Leu Leu Pro Arg AlaPro Leu 35 40 45 Ala Arg Ala Arg Ala Arg Ala Ala Leu Ala Arg Ala Ala LeuAla Pro 50 55 60 Arg Leu Pro His Asn Leu Ser Leu Glu Leu Val Val Ala AlaPro Pro 65 70 75 80 Ala Arg Asp Pro Ala Ser Leu Thr Arg Gly Leu Cys GlnAla Leu Val 85 90 95 Pro Pro Gly Val Ala Ala Leu Leu Ala Phe Pro Glu AlaArg Pro Glu 100 105 110 Leu Leu Gln Leu His Phe Leu Ala Ala Ala Thr GluThr Pro Val Leu 115 120 125 Ser Leu Leu Arg Arg Glu Ala Arg Ala Pro LeuGly Ala Pro Asn Pro 130 135 140 Phe His Leu Gln Leu His Trp Ala Ser ProLeu Glu Thr Leu Leu Asp 145 150 155 160 Val Leu Val Ala Val Leu Gln AlaHis Ala Trp Glu Asp Val Gly Leu 165 170 175 Ala Leu Cys Arg Thr Gln AspPro Gly Gly Leu Val Ala Leu Trp Thr 180 185 190 Ser Arg Ala Gly Arg ProPro Gln Leu Val Leu Asp Leu Ser Arg Arg 195 200 205 Asp Thr Gly Asp AlaGly Leu Arg Ala Arg Leu Ala Pro Met Ala Ala 210 215 220 Pro Val Gly GlyGlu Ala Pro Val Pro Ala Ala Val Leu Leu Gly Cys 225 230 235 240 Asp IleAla Arg Ala Arg Arg Val Leu Glu Ala Val Pro Pro Gly Pro 245 250 255 HisTrp Leu Leu Gly Thr Pro Leu Pro Pro Lys Ala Leu Pro Thr Ala 260 265 270Gly Leu Pro Pro Gly Leu Leu Ala Leu Gly Glu Val Ala Arg Pro Pro 275 280285 Leu Glu Ala Ala Ile His Asp Ile Val Gln Leu Val Ala Arg Ala Leu 290295 300 Gly Ser Ala Ala Gln Val Gln Pro Lys Arg Ala Leu Leu Pro Ala Pro305 310 315 320 Val Asn Cys Gly Asp Leu Gln Pro Ala Gly Pro Glu Ser ProGly Arg 325 330 335 Phe Leu Ala Arg Phe Leu Ala Asn Thr Ser Phe Gln GlyArg Thr Gly 340 345 350 Pro Val Trp Val Thr Gly Ser Ser Pro Asp Glu AspGly Gln Cys Pro 355 360 365 Ala Gly Gln Leu Cys Leu Asp Pro Gly Thr AsnAsp Ser Ala Thr Leu 370 375 380 Asp Ala Leu Phe Ala Ala Leu Ala Asn GlySer Ala Pro Arg Ala Leu 385 390 395 400 Arg Lys Cys Cys Tyr Gly Tyr CysIle Asp Leu Leu Glu Arg Leu Ala 405 410 415 Glu Asp Thr Pro Phe Asp PheGlu Leu Tyr Leu Val Gly Asp Gly Lys 420 425 430 Tyr Gly Ala Leu Arg AspGly Arg Trp Thr Gly Leu Val Gly Asp Leu 435 440 445 Leu Ala Gly Arg AlaHis Met Ala Val Thr Ser Phe Ser Ile Asn Ser 450 455 460 Ala Arg Ser GlnVal Val Asp Phe Thr Ser Pro Phe Phe Ser Thr Ser 465 470 475 480 Leu GlyIle Met Val Arg Ala Arg Asp Thr Ala Ser Pro Ile Gly Ala 485 490 495 PheMet Trp Pro Leu His Trp Ser Thr Trp Leu Gly Val Phe Ala Ala 500 505 510Leu His Leu Thr Ala Leu Phe Leu Thr Val Tyr Glu Trp Arg Ser Pro 515 520525 Tyr Gly Leu Thr Pro Arg Gly Arg Asn Arg Ser Thr Val Phe Ser Tyr 530535 540 Ser Ser Ala Leu Asn Leu Cys Tyr Ala Ile Leu Phe Arg Arg Thr Val545 550 555 560 Ser Ser Lys Thr Pro Lys Cys Pro Thr Gly Arg Leu Leu MetAsn Leu 565 570 575 Trp Ala Ile Phe Cys Leu Leu Val Leu Ser Ser Tyr ThrAla Asn Leu 580 585 590 Ala Ala Val Met Val Gly Asp Lys Thr Phe Glu GluLeu Ser Gly Ile 595 600 605 His Asp Pro Lys Leu His His Pro Ala Gln GlyPhe Arg Phe Gly Thr 610 615 620 Val Trp Glu Ser Ser Ala Glu Ala Tyr IleLys Lys Ser Phe Pro Asp 625 630 635 640 Met His Ala His Met Arg Arg HisSer Ala Pro Thr Thr Pro Arg Gly 645 650 655 Val Ala Met Leu Thr Ser AspPro Pro Lys Leu Asn Ala Phe Ile Met 660 665 670 Asp Lys Ser Leu Leu AspTyr Glu Val Ser Ile Asp Ala Asp Cys Lys 675 680 685 Leu Leu Thr Val GlyLys Pro Phe Ala Ile Glu Gly Tyr Gly Ile Gly 690 695 700 Leu Pro Gln AsnSer Pro Leu Thr Ser Asn Leu Ser Glu Phe Ile Ser 705 710 715 720 Arg TyrLys Ser Ser Gly Phe Ile Asp Leu Leu His Asp Lys Trp Tyr 725 730 735 LysMet Val Pro Cys Gly Lys Arg Val Phe Ala Val Thr Glu Thr Leu 740 745 750Gln Met Ser Ile Tyr His Phe Ala Gly Leu Phe Val Leu Leu Cys Leu 755 760765 Gly Leu Gly Ser Ala Leu Leu Ser Ser Leu Gly Glu His Ala Phe Phe 770775 780 Arg Leu Ala Leu Pro Arg Ile Arg Lys Gly Ser Arg Leu Gln Tyr Trp785 790 795 800 Leu His Thr Ser Gln Lys Ile His Arg Ala Leu Asn Thr GluPro Pro 805 810 815 Glu Gly Ser Lys Glu Glu Thr Ala Glu Ala Glu Pro SerGly Pro Glu 820 825 830 Val Glu Gln Gln Gln Gln Gln Gln Asp Gln Pro ThrAla Pro Glu Gly 835 840 845 Trp Lys Arg Ala Arg Arg Ala Val Asp Lys GluArg Arg Val Arg Phe 850 855 860 Leu Leu Glu Pro Ala Val Val Val Ala ProGlu Ala Asp Ala Glu Ala 865 870 875 880 Glu Ala Ala Pro Arg Glu Gly ProVal Trp Leu Cys Ser Tyr Gly Arg 885 890 895 Pro Pro Ala Ala Arg Pro ThrGly Ala Pro Gln Pro Gly Glu Leu Gln 900 905 910 Glu Leu Glu Arg Arg IleGlu Val Ala Arg Glu Arg Leu Arg Gln Ala 915 920 925 Leu Val Arg Arg GlyGln Leu Leu Ala Gln Leu Gly Asp Ser Ala Arg 930 935 940 His Arg Pro ArgArg Leu Leu Gln Ala Arg Ala Ala Pro Ala Glu Ala 945 950 955 960 Pro ProHis Ser Gly Arg Pro Gly Ser Gln Glu 965 970 7 3132 DNA Homo sapiens CDS(1)..(3129) 7 atg gag ttt gtg cgg gcg ctg tgg ctg ggc ctg gcg ctg gcgctg ggg 48 Met Glu Phe Val Arg Ala Leu Trp Leu Gly Leu Ala Leu Ala LeuGly 1 5 10 15 ccg ggg tcc gcg ggg ggc cac cct cag ccg tgc ggc gtc ctggcg cgc 96 Pro Gly Ser Ala Gly Gly His Pro Gln Pro Cys Gly Val Leu AlaArg 20 25 30 ctc ggg ggc tcc gtg cgc ctg ggc gcc ctc ctg ccc cgc gcg cctctc 144 Leu Gly Gly Ser Val Arg Leu Gly Ala Leu Leu Pro Arg Ala Pro Leu35 40 45 gcc cgc gcc cgc gcc cgc gcc gcc ctg gcc cgg gcc gcc ctg gcg ccg192 Ala Arg Ala Arg Ala Arg Ala Ala Leu Ala Arg Ala Ala Leu Ala Pro 5055 60 cgg ctg ccg cac aac ctg agc ttg gag ctg gtg gtc gcc gcg ccc ccc240 Arg Leu Pro His Asn Leu Ser Leu Glu Leu Val Val Ala Ala Pro Pro 6570 75 80 gcc cgc gac ccc gcc tcg ctg acc cgc ggc ctg tgc cag gcg ctg gtg288 Ala Arg Asp Pro Ala Ser Leu Thr Arg Gly Leu Cys Gln Ala Leu Val 8590 95 cct ccg ggc gtg gcg gcc ctg ctc gcc ttt ccc gag gct cgg ccc gag336 Pro Pro Gly Val Ala Ala Leu Leu Ala Phe Pro Glu Ala Arg Pro Glu 100105 110 ctg ctg cag ctg cac ttc ctg gcg gcg gcc acc gag acc ccc gtg ctc384 Leu Leu Gln Leu His Phe Leu Ala Ala Ala Thr Glu Thr Pro Val Leu 115120 125 agc ctg ctg cgg cgg gag gcg cgc gcg ccc ctc gga gcc ccg aac cca432 Ser Leu Leu Arg Arg Glu Ala Arg Ala Pro Leu Gly Ala Pro Asn Pro 130135 140 ttc cac ctg cag ctg cac tgg gcc agc ccc ctg gag acg ctg ctg gat480 Phe His Leu Gln Leu His Trp Ala Ser Pro Leu Glu Thr Leu Leu Asp 145150 155 160 gtg ctg gtg gcg gtg ctg cag gcg cac gcc tgg gaa gac gtc ggcctg 528 Val Leu Val Ala Val Leu Gln Ala His Ala Trp Glu Asp Val Gly Leu165 170 175 gcc ctg tgc cgc act cag gac ccc ggc ggc ctg gtg gcc ctc tggaca 576 Ala Leu Cys Arg Thr Gln Asp Pro Gly Gly Leu Val Ala Leu Trp Thr180 185 190 agc cgg gct ggc cgg ccc cca cag ctg gtc ctg gac cta agc cggcgg 624 Ser Arg Ala Gly Arg Pro Pro Gln Leu Val Leu Asp Leu Ser Arg Arg195 200 205 gac acg gga gat gca gga ctg cgg gca cgc ctg gcc ccg atg gcggcg 672 Asp Thr Gly Asp Ala Gly Leu Arg Ala Arg Leu Ala Pro Met Ala Ala210 215 220 cca gtg ggg ggt gaa gca ccg gta ccc gcg gcg gtc ctc ctc ggctgt 720 Pro Val Gly Gly Glu Ala Pro Val Pro Ala Ala Val Leu Leu Gly Cys225 230 235 240 gac atc gcc cgt gcc cgt cgg gtg ctg gag gcc gta cct cccggc ccc 768 Asp Ile Ala Arg Ala Arg Arg Val Leu Glu Ala Val Pro Pro GlyPro 245 250 255 cac tgg ctg ttg ggg aca cca ctg ccg ccc aag gcc ctg cccacc gcg 816 His Trp Leu Leu Gly Thr Pro Leu Pro Pro Lys Ala Leu Pro ThrAla 260 265 270 ggg ctg cca cca ggg ctg ctg gcg ctg ggc gag gtg gca cgaccc ccg 864 Gly Leu Pro Pro Gly Leu Leu Ala Leu Gly Glu Val Ala Arg ProPro 275 280 285 ctg gag gcc gcc atc cat gac att gtg caa ctg gtg gcc cgggcg ctg 912 Leu Glu Ala Ala Ile His Asp Ile Val Gln Leu Val Ala Arg AlaLeu 290 295 300 ggc agt gcg gcc cag gtg cag ccg aag cga gcc ctc ctc cccgcc ccg 960 Gly Ser Ala Ala Gln Val Gln Pro Lys Arg Ala Leu Leu Pro AlaPro 305 310 315 320 gtc aac tgc ggg gac ctg cag ccg gcc ggg ccc gag tccccg ggg cgc 1008 Val Asn Cys Gly Asp Leu Gln Pro Ala Gly Pro Glu Ser ProGly Arg 325 330 335 ttc ttg gca cgg ttc ctg gcc aac acg tcc ttc cag ggccgc acg ggc 1056 Phe Leu Ala Arg Phe Leu Ala Asn Thr Ser Phe Gln Gly ArgThr Gly 340 345 350 ccc gtg tgg gtg aca ggc agc tcc cag gta cac atg tctcgg cac ttt 1104 Pro Val Trp Val Thr Gly Ser Ser Gln Val His Met Ser ArgHis Phe 355 360 365 aag gtg tgg agc ctt cgc cgg gac cca cgg ggc gcc ccggcc tgg gcc 1152 Lys Val Trp Ser Leu Arg Arg Asp Pro Arg Gly Ala Pro AlaTrp Ala 370 375 380 acg gtg ggc agc tgg cgg gac ggc cag ctg gac ttg gaaccg gga ggt 1200 Thr Val Gly Ser Trp Arg Asp Gly Gln Leu Asp Leu Glu ProGly Gly 385 390 395 400 gcc tct gca cgg ccc ccg ccc cca cag ggt gcc caggtc tgg ccc aag 1248 Ala Ser Ala Arg Pro Pro Pro Pro Gln Gly Ala Gln ValTrp Pro Lys 405 410 415 ctg cgt gtg gta acg ctg ttg gaa cac cca ttt gtgttt gcc cgt gat 1296 Leu Arg Val Val Thr Leu Leu Glu His Pro Phe Val PheAla Arg Asp 420 425 430 cca gac gaa gac ggg cag tgc cca gcg ggg cag ctgtgc ctg gac cct 1344 Pro Asp Glu Asp Gly Gln Cys Pro Ala Gly Gln Leu CysLeu Asp Pro 435 440 445 ggc acc aac gac tcg gcc acc ctg gac gca ctg ttcgcc gcg ctg gcc 1392 Gly Thr Asn Asp Ser Ala Thr Leu Asp Ala Leu Phe AlaAla Leu Ala 450 455 460 aac ggc tca gcg ccc cgt gcc ctg cgc aag tgc tgctac ggc tac tgc 1440 Asn Gly Ser Ala Pro Arg Ala Leu Arg Lys Cys Cys TyrGly Tyr Cys 465 470 475 480 att gac ctg ctg gag cgg ctg gcg gag gac acgccc ttc gac ttc gag 1488 Ile Asp Leu Leu Glu Arg Leu Ala Glu Asp Thr ProPhe Asp Phe Glu 485 490 495 ctg tac ctc gtg ggt gac ggc aag tac ggc gccctg cgg gac ggc cgc 1536 Leu Tyr Leu Val Gly Asp Gly Lys Tyr Gly Ala LeuArg Asp Gly Arg 500 505 510 tgg acc ggc ctg gtc ggg gac ctg ctg gcc ggccgg gcc cac atg gcg 1584 Trp Thr Gly Leu Val Gly Asp Leu Leu Ala Gly ArgAla His Met Ala 515 520 525 gtc acc agc ttc agt atc aac tcc gcc cgc tcacag gtg gtg gac ttc 1632 Val Thr Ser Phe Ser Ile Asn Ser Ala Arg Ser GlnVal Val Asp Phe 530 535 540 acc agc ccc ttc ttc tcc acc agc ctg ggc atcatg gtg cgg gca cgg 1680 Thr Ser Pro Phe Phe Ser Thr Ser Leu Gly Ile MetVal Arg Ala Arg 545 550 555 560 gac acg gcc tca ccc atc ggt gcc ttt atgtgg ccc ctg cac tgg tcc 1728 Asp Thr Ala Ser Pro Ile Gly Ala Phe Met TrpPro Leu His Trp Ser 565 570 575 acg tgg ctg ggc gtc ttt gcg gcc ctg cacctc acc gcg ctc ttc ctc 1776 Thr Trp Leu Gly Val Phe Ala Ala Leu His LeuThr Ala Leu Phe Leu 580 585 590 acc gtg tac gag tgg cgt agc ccc tac ggcctc acg cca cgt ggc cgc 1824 Thr Val Tyr Glu Trp Arg Ser Pro Tyr Gly LeuThr Pro Arg Gly Arg 595 600 605 aac cgc agc acc gtc ttc tcc tac tcc tcagcc ctc aac ctg tgc tac 1872 Asn Arg Ser Thr Val Phe Ser Tyr Ser Ser AlaLeu Asn Leu Cys Tyr 610 615 620 gcc atc ctc ttc aga cgc acc gtg tcc agcaag acg ccc aag tgc ccc 1920 Ala Ile Leu Phe Arg Arg Thr Val Ser Ser LysThr Pro Lys Cys Pro 625 630 635 640 acg ggc cgc ctg ctc atg aac ctc tgggcc atc ttc tgc ctg ctg gtg 1968 Thr Gly Arg Leu Leu Met Asn Leu Trp AlaIle Phe Cys Leu Leu Val 645 650 655 ctg tcc agc tac acg gcc aac ctg gctgcc gtc atg gtc ggg gac aag 2016 Leu Ser Ser Tyr Thr Ala Asn Leu Ala AlaVal Met Val Gly Asp Lys 660 665 670 acc ttc gag gag ctg tcg ggg atc cacgac ccc aag ctg cac cac ccg 2064 Thr Phe Glu Glu Leu Ser Gly Ile His AspPro Lys Leu His His Pro 675 680 685 gcg cag ggc ttc cgc ttc ggc acc gtgtgg gag agc agc gcc gag gcg 2112 Ala Gln Gly Phe Arg Phe Gly Thr Val TrpGlu Ser Ser Ala Glu Ala 690 695 700 tac atc aag aag agc ttc ccc gac atgcac gca cac atg cgg cgc cac 2160 Tyr Ile Lys Lys Ser Phe Pro Asp Met HisAla His Met Arg Arg His 705 710 715 720 agc gcg ccc acc acg ccc cgc ggcgtc gcc atg ctc acg agc gac ccc 2208 Ser Ala Pro Thr Thr Pro Arg Gly ValAla Met Leu Thr Ser Asp Pro 725 730 735 ccc aag ctc aac gcc ttc atc atggac aag tcg ctc ctg gac tac gag 2256 Pro Lys Leu Asn Ala Phe Ile Met AspLys Ser Leu Leu Asp Tyr Glu 740 745 750 gtc tcc atc gac gcc gac tgc aaactg ctg acc gtg gga aag ccc ttc 2304 Val Ser Ile Asp Ala Asp Cys Lys LeuLeu Thr Val Gly Lys Pro Phe 755 760 765 gcc att gag ggc tat ggg atc ggactg ccc cag aac tcg ccg ctc acc 2352 Ala Ile Glu Gly Tyr Gly Ile Gly LeuPro Gln Asn Ser Pro Leu Thr 770 775 780 tcc aac ctg tcc gag ttc atc agccgc tac aag tcc tcc ggc ttc atc 2400 Ser Asn Leu Ser Glu Phe Ile Ser ArgTyr Lys Ser Ser Gly Phe Ile 785 790 795 800 gac ctg ctc cac gac aag tggtac aag atg gtg cct tgc ggc aag cgg 2448 Asp Leu Leu His Asp Lys Trp TyrLys Met Val Pro Cys Gly Lys Arg 805 810 815 gtc ttt gcg gtt aca gag accctg cag atg agc atc tac cac ttc gcg 2496 Val Phe Ala Val Thr Glu Thr LeuGln Met Ser Ile Tyr His Phe Ala 820 825 830 ggc ctc ttc gtg ttg ctg tgcctg ggc ctg ggc agc gct ctg ctc agc 2544 Gly Leu Phe Val Leu Leu Cys LeuGly Leu Gly Ser Ala Leu Leu Ser 835 840 845 tcg ctg ggc gag cac gcc ttcttc cgc ctg gcg ctg ccg cgc atc cgc 2592 Ser Leu Gly Glu His Ala Phe PheArg Leu Ala Leu Pro Arg Ile Arg 850 855 860 aag ggg agc agg ctg cag tactgg ctg cac acc agc cag aaa atc cac 2640 Lys Gly Ser Arg Leu Gln Tyr TrpLeu His Thr Ser Gln Lys Ile His 865 870 875 880 cgc gcc ctc aac acg gagcca cca gag ggg tcg aag gag gag acg gca 2688 Arg Ala Leu Asn Thr Glu ProPro Glu Gly Ser Lys Glu Glu Thr Ala 885 890 895 gag gcg gag ccc agc ggcccc gag gtg gag cag cag cag cag cag cag 2736 Glu Ala Glu Pro Ser Gly ProGlu Val Glu Gln Gln Gln Gln Gln Gln 900 905 910 gac cag cca acg gct ccggag ggc tgg aaa cgg gcg cgc cgg gcc gtg 2784 Asp Gln Pro Thr Ala Pro GluGly Trp Lys Arg Ala Arg Arg Ala Val 915 920 925 gac aag gag cgc cgc gtgcgc ttc ctg ctg gag ccc gcc gtg gtt gtg 2832 Asp Lys Glu Arg Arg Val ArgPhe Leu Leu Glu Pro Ala Val Val Val 930 935 940 gca ccc gaa gcg gac gcggag gcg gag gct gcg ccg cga gag ggc ccc 2880 Ala Pro Glu Ala Asp Ala GluAla Glu Ala Ala Pro Arg Glu Gly Pro 945 950 955 960 gtc tgg ctg tgc tcctac ggc cgc ccg ccc gcc gca agg ccc acg ggg 2928 Val Trp Leu Cys Ser TyrGly Arg Pro Pro Ala Ala Arg Pro Thr Gly 965 970 975 gcc ccc cag ccc ggggag ctg cag gag ctg gag cgc cgc atc gaa gtc 2976 Ala Pro Gln Pro Gly GluLeu Gln Glu Leu Glu Arg Arg Ile Glu Val 980 985 990 gcg cgt gag cgg ctccgc cag gcc ctg gtg cgg cgc ggc cag ctc ctg 3024 Ala Arg Glu Arg Leu ArgGln Ala Leu Val Arg Arg Gly Gln Leu Leu 995 1000 1005 gca cag ctc ggggac agc gca cgt cac cgg cct cgg cgc ttg ctt cag 3072 Ala Gln Leu Gly AspSer Ala Arg His Arg Pro Arg Arg Leu Leu Gln 1010 1015 1020 gcc aga gcggcc ccc gcg gag gcc cca cca cac tct ggc cga ccg ggg 3120 Ala Arg Ala AlaPro Ala Glu Ala Pro Pro His Ser Gly Arg Pro Gly 1025 1030 1035 1040 agccag gaa tga 3132 Ser Gln Glu 8 1043 PRT Homo sapiens 8 Met Glu Phe ValArg Ala Leu Trp Leu Gly Leu Ala Leu Ala Leu Gly 1 5 10 15 Pro Gly SerAla Gly Gly His Pro Gln Pro Cys Gly Val Leu Ala Arg 20 25 30 Leu Gly GlySer Val Arg Leu Gly Ala Leu Leu Pro Arg Ala Pro Leu 35 40 45 Ala Arg AlaArg Ala Arg Ala Ala Leu Ala Arg Ala Ala Leu Ala Pro 50 55 60 Arg Leu ProHis Asn Leu Ser Leu Glu Leu Val Val Ala Ala Pro Pro 65 70 75 80 Ala ArgAsp Pro Ala Ser Leu Thr Arg Gly Leu Cys Gln Ala Leu Val 85 90 95 Pro ProGly Val Ala Ala Leu Leu Ala Phe Pro Glu Ala Arg Pro Glu 100 105 110 LeuLeu Gln Leu His Phe Leu Ala Ala Ala Thr Glu Thr Pro Val Leu 115 120 125Ser Leu Leu Arg Arg Glu Ala Arg Ala Pro Leu Gly Ala Pro Asn Pro 130 135140 Phe His Leu Gln Leu His Trp Ala Ser Pro Leu Glu Thr Leu Leu Asp 145150 155 160 Val Leu Val Ala Val Leu Gln Ala His Ala Trp Glu Asp Val GlyLeu 165 170 175 Ala Leu Cys Arg Thr Gln Asp Pro Gly Gly Leu Val Ala LeuTrp Thr 180 185 190 Ser Arg Ala Gly Arg Pro Pro Gln Leu Val Leu Asp LeuSer Arg Arg 195 200 205 Asp Thr Gly Asp Ala Gly Leu Arg Ala Arg Leu AlaPro Met Ala Ala 210 215 220 Pro Val Gly Gly Glu Ala Pro Val Pro Ala AlaVal Leu Leu Gly Cys 225 230 235 240 Asp Ile Ala Arg Ala Arg Arg Val LeuGlu Ala Val Pro Pro Gly Pro 245 250 255 His Trp Leu Leu Gly Thr Pro LeuPro Pro Lys Ala Leu Pro Thr Ala 260 265 270 Gly Leu Pro Pro Gly Leu LeuAla Leu Gly Glu Val Ala Arg Pro Pro 275 280 285 Leu Glu Ala Ala Ile HisAsp Ile Val Gln Leu Val Ala Arg Ala Leu 290 295 300 Gly Ser Ala Ala GlnVal Gln Pro Lys Arg Ala Leu Leu Pro Ala Pro 305 310 315 320 Val Asn CysGly Asp Leu Gln Pro Ala Gly Pro Glu Ser Pro Gly Arg 325 330 335 Phe LeuAla Arg Phe Leu Ala Asn Thr Ser Phe Gln Gly Arg Thr Gly 340 345 350 ProVal Trp Val Thr Gly Ser Ser Gln Val His Met Ser Arg His Phe 355 360 365Lys Val Trp Ser Leu Arg Arg Asp Pro Arg Gly Ala Pro Ala Trp Ala 370 375380 Thr Val Gly Ser Trp Arg Asp Gly Gln Leu Asp Leu Glu Pro Gly Gly 385390 395 400 Ala Ser Ala Arg Pro Pro Pro Pro Gln Gly Ala Gln Val Trp ProLys 405 410 415 Leu Arg Val Val Thr Leu Leu Glu His Pro Phe Val Phe AlaArg Asp 420 425 430 Pro Asp Glu Asp Gly Gln Cys Pro Ala Gly Gln Leu CysLeu Asp Pro 435 440 445 Gly Thr Asn Asp Ser Ala Thr Leu Asp Ala Leu PheAla Ala Leu Ala 450 455 460 Asn Gly Ser Ala Pro Arg Ala Leu Arg Lys CysCys Tyr Gly Tyr Cys 465 470 475 480 Ile Asp Leu Leu Glu Arg Leu Ala GluAsp Thr Pro Phe Asp Phe Glu 485 490 495 Leu Tyr Leu Val Gly Asp Gly LysTyr Gly Ala Leu Arg Asp Gly Arg 500 505 510 Trp Thr Gly Leu Val Gly AspLeu Leu Ala Gly Arg Ala His Met Ala 515 520 525 Val Thr Ser Phe Ser IleAsn Ser Ala Arg Ser Gln Val Val Asp Phe 530 535 540 Thr Ser Pro Phe PheSer Thr Ser Leu Gly Ile Met Val Arg Ala Arg 545 550 555 560 Asp Thr AlaSer Pro Ile Gly Ala Phe Met Trp Pro Leu His Trp Ser 565 570 575 Thr TrpLeu Gly Val Phe Ala Ala Leu His Leu Thr Ala Leu Phe Leu 580 585 590 ThrVal Tyr Glu Trp Arg Ser Pro Tyr Gly Leu Thr Pro Arg Gly Arg 595 600 605Asn Arg Ser Thr Val Phe Ser Tyr Ser Ser Ala Leu Asn Leu Cys Tyr 610 615620 Ala Ile Leu Phe Arg Arg Thr Val Ser Ser Lys Thr Pro Lys Cys Pro 625630 635 640 Thr Gly Arg Leu Leu Met Asn Leu Trp Ala Ile Phe Cys Leu LeuVal 645 650 655 Leu Ser Ser Tyr Thr Ala Asn Leu Ala Ala Val Met Val GlyAsp Lys 660 665 670 Thr Phe Glu Glu Leu Ser Gly Ile His Asp Pro Lys LeuHis His Pro 675 680 685 Ala Gln Gly Phe Arg Phe Gly Thr Val Trp Glu SerSer Ala Glu Ala 690 695 700 Tyr Ile Lys Lys Ser Phe Pro Asp Met His AlaHis Met Arg Arg His 705 710 715 720 Ser Ala Pro Thr Thr Pro Arg Gly ValAla Met Leu Thr Ser Asp Pro 725 730 735 Pro Lys Leu Asn Ala Phe Ile MetAsp Lys Ser Leu Leu Asp Tyr Glu 740 745 750 Val Ser Ile Asp Ala Asp CysLys Leu Leu Thr Val Gly Lys Pro Phe 755 760 765 Ala Ile Glu Gly Tyr GlyIle Gly Leu Pro Gln Asn Ser Pro Leu Thr 770 775 780 Ser Asn Leu Ser GluPhe Ile Ser Arg Tyr Lys Ser Ser Gly Phe Ile 785 790 795 800 Asp Leu LeuHis Asp Lys Trp Tyr Lys Met Val Pro Cys Gly Lys Arg 805 810 815 Val PheAla Val Thr Glu Thr Leu Gln Met Ser Ile Tyr His Phe Ala 820 825 830 GlyLeu Phe Val Leu Leu Cys Leu Gly Leu Gly Ser Ala Leu Leu Ser 835 840 845Ser Leu Gly Glu His Ala Phe Phe Arg Leu Ala Leu Pro Arg Ile Arg 850 855860 Lys Gly Ser Arg Leu Gln Tyr Trp Leu His Thr Ser Gln Lys Ile His 865870 875 880 Arg Ala Leu Asn Thr Glu Pro Pro Glu Gly Ser Lys Glu Glu ThrAla 885 890 895 Glu Ala Glu Pro Ser Gly Pro Glu Val Glu Gln Gln Gln GlnGln Gln 900 905 910 Asp Gln Pro Thr Ala Pro Glu Gly Trp Lys Arg Ala ArgArg Ala Val 915 920 925 Asp Lys Glu Arg Arg Val Arg Phe Leu Leu Glu ProAla Val Val Val 930 935 940 Ala Pro Glu Ala Asp Ala Glu Ala Glu Ala AlaPro Arg Glu Gly Pro 945 950 955 960 Val Trp Leu Cys Ser Tyr Gly Arg ProPro Ala Ala Arg Pro Thr Gly 965 970 975 Ala Pro Gln Pro Gly Glu Leu GlnGlu Leu Glu Arg Arg Ile Glu Val 980 985 990 Ala Arg Glu Arg Leu Arg GlnAla Leu Val Arg Arg Gly Gln Leu Leu 995 1000 1005 Ala Gln Leu Gly AspSer Ala Arg His Arg Pro Arg Arg Leu Leu Gln 1010 1015 1020 Ala Arg AlaAla Pro Ala Glu Ala Pro Pro His Ser Gly Arg Pro Gly 1025 1030 1035 1040Ser Gln Glu 9 1110 DNA Homo sapiens CDS (163)..(990) 9 acgcgttactcctaccaggt tgtagcatgc atctttttga gagagcagct gggatcgagt 60 atactcttgacttaaatatg tttgtttata aagacaaatg gagaaatcaa tttttttccc 120 tgaattcttaggagcacttt agtgaataaa gaacctgaca gt atg ctg gcc cac 174 Met Leu Ala His1 atg ttt aag gac aaa ggt gtc tgg gga aat aag caa gat cat aga gga 222Met Phe Lys Asp Lys Gly Val Trp Gly Asn Lys Gln Asp His Arg Gly 5 10 1520 gct ttc tta att gac cga agt cct gag tac ttc gaa ccc att ttg aac 270Ala Phe Leu Ile Asp Arg Ser Pro Glu Tyr Phe Glu Pro Ile Leu Asn 25 30 35tac ttg cgt cat gga cag ctc att gta aat gat ggc att aat tta ttg 318 TyrLeu Arg His Gly Gln Leu Ile Val Asn Asp Gly Ile Asn Leu Leu 40 45 50 ggtgtg tta gaa gaa gca aga ttt ttt ggt att gac tca ttg att gaa 366 Gly ValLeu Glu Glu Ala Arg Phe Phe Gly Ile Asp Ser Leu Ile Glu 55 60 65 cac ctagaa gtg gca ata aag aat tct caa cca ccg gag gat cat tca 414 His Leu GluVal Ala Ile Lys Asn Ser Gln Pro Pro Glu Asp His Ser 70 75 80 cca ata tcccga aag gaa ttt gtc cga ttt ttg cta gca act cca acc 462 Pro Ile Ser ArgLys Glu Phe Val Arg Phe Leu Leu Ala Thr Pro Thr 85 90 95 100 aag tca gaactg cga tgc cag ggt ttg aac ttc agt ggt gct gat ctt 510 Lys Ser Glu LeuArg Cys Gln Gly Leu Asn Phe Ser Gly Ala Asp Leu 105 110 115 tct cgt ttggac ctt cga tac att aac ttc aaa atg gcc aat tta agc 558 Ser Arg Leu AspLeu Arg Tyr Ile Asn Phe Lys Met Ala Asn Leu Ser 120 125 130 cgc tgt aatctt gca cat gca aat ctt tgc tgt gca aat ctt gaa cga 606 Arg Cys Asn LeuAla His Ala Asn Leu Cys Cys Ala Asn Leu Glu Arg 135 140 145 gct gat ctctct gga tca gtg ctt gac tgt gcg aat ctc cag gga gtc 654 Ala Asp Leu SerGly Ser Val Leu Asp Cys Ala Asn Leu Gln Gly Val 150 155 160 aag atg ctctgt tct aat gca gaa gga gca tcc ctg aaa ctg tgt aat 702 Lys Met Leu CysSer Asn Ala Glu Gly Ala Ser Leu Lys Leu Cys Asn 165 170 175 180 ttt gaggat cct tct ggt ctt aaa gcc aat tta gaa ggt gct aat ctg 750 Phe Glu AspPro Ser Gly Leu Lys Ala Asn Leu Glu Gly Ala Asn Leu 185 190 195 aaa ggtgtg gat atg gaa gga agt cag atg aca gga att aac ctg aga 798 Lys Gly ValAsp Met Glu Gly Ser Gln Met Thr Gly Ile Asn Leu Arg 200 205 210 gtg gctacc tta aaa aat gca aag ttg aag aac tgt aac ctc aga gga 846 Val Ala ThrLeu Lys Asn Ala Lys Leu Lys Asn Cys Asn Leu Arg Gly 215 220 225 gca actctg gca gga act gat tta gag aat tgt gat ctg tct ggg tgt 894 Ala Thr LeuAla Gly Thr Asp Leu Glu Asn Cys Asp Leu Ser Gly Cys 230 235 240 gat cttcaa gaa gcc aac ctg aga ggg tcc aac gtg aag gga gct ata 942 Asp Leu GlnGlu Ala Asn Leu Arg Gly Ser Asn Val Lys Gly Ala Ile 245 250 255 260 tttgaa gag atg ctg aca cca cta cac atg tca caa agt gtc aga tga 990 Phe GluGlu Met Leu Thr Pro Leu His Met Ser Gln Ser Val Arg 265 270 275gaattttagg ggctggagga agatgtaaaa gatgaaaatg ttttccttat cacttttctt 1050tctccaccca ctcagttgtc tagaagaaat aacactgtaa ggaaatttaa aaaaaaaaaa 111010 275 PRT Homo sapiens 10 Met Leu Ala His Met Phe Lys Asp Lys Gly ValTrp Gly Asn Lys Gln 1 5 10 15 Asp His Arg Gly Ala Phe Leu Ile Asp ArgSer Pro Glu Tyr Phe Glu 20 25 30 Pro Ile Leu Asn Tyr Leu Arg His Gly GlnLeu Ile Val Asn Asp Gly 35 40 45 Ile Asn Leu Leu Gly Val Leu Glu Glu AlaArg Phe Phe Gly Ile Asp 50 55 60 Ser Leu Ile Glu His Leu Glu Val Ala IleLys Asn Ser Gln Pro Pro 65 70 75 80 Glu Asp His Ser Pro Ile Ser Arg LysGlu Phe Val Arg Phe Leu Leu 85 90 95 Ala Thr Pro Thr Lys Ser Glu Leu ArgCys Gln Gly Leu Asn Phe Ser 100 105 110 Gly Ala Asp Leu Ser Arg Leu AspLeu Arg Tyr Ile Asn Phe Lys Met 115 120 125 Ala Asn Leu Ser Arg Cys AsnLeu Ala His Ala Asn Leu Cys Cys Ala 130 135 140 Asn Leu Glu Arg Ala AspLeu Ser Gly Ser Val Leu Asp Cys Ala Asn 145 150 155 160 Leu Gln Gly ValLys Met Leu Cys Ser Asn Ala Glu Gly Ala Ser Leu 165 170 175 Lys Leu CysAsn Phe Glu Asp Pro Ser Gly Leu Lys Ala Asn Leu Glu 180 185 190 Gly AlaAsn Leu Lys Gly Val Asp Met Glu Gly Ser Gln Met Thr Gly 195 200 205 IleAsn Leu Arg Val Ala Thr Leu Lys Asn Ala Lys Leu Lys Asn Cys 210 215 220Asn Leu Arg Gly Ala Thr Leu Ala Gly Thr Asp Leu Glu Asn Cys Asp 225 230235 240 Leu Ser Gly Cys Asp Leu Gln Glu Ala Asn Leu Arg Gly Ser Asn Val245 250 255 Lys Gly Ala Ile Phe Glu Glu Met Leu Thr Pro Leu His Met SerGln 260 265 270 Ser Val Arg 275 11 926 DNA Homo sapiens CDS (33)..(887)11 tttccagggt tctagcctgt tcatctagcc cc atg atg gct gtg gac atc gag 53Met Met Ala Val Asp Ile Glu 1 5 tac aga tac aac tgc atg gct cct tcc ttgcgc caa gag agg ttt gcc 101 Tyr Arg Tyr Asn Cys Met Ala Pro Ser Leu ArgGln Glu Arg Phe Ala 10 15 20 ttt aag atc tca cca aag ccc agc aaa cca ctgagg cct tgt att cag 149 Phe Lys Ile Ser Pro Lys Pro Ser Lys Pro Leu ArgPro Cys Ile Gln 25 30 35 ctg agc agc aag aat gaa gcc agt gga atg gtg gccccg gct gtc cag 197 Leu Ser Ser Lys Asn Glu Ala Ser Gly Met Val Ala ProAla Val Gln 40 45 50 55 gag aag aag gtg aaa aag cgg gtg tcc ttc gca gacaac cag ggg ctg 245 Glu Lys Lys Val Lys Lys Arg Val Ser Phe Ala Asp AsnGln Gly Leu 60 65 70 gcc ctg aca atg gtc aaa gtg ttc tcg gaa ttc gat gacccg cta gat 293 Ala Leu Thr Met Val Lys Val Phe Ser Glu Phe Asp Asp ProLeu Asp 75 80 85 atg cca ttc aac atc acc gag ctc cta gac aac att gtg agcttg acg 341 Met Pro Phe Asn Ile Thr Glu Leu Leu Asp Asn Ile Val Ser LeuThr 90 95 100 aca gca gag agc gag agc ttt gtt ctg gat ttt tcc cag ccctct gca 389 Thr Ala Glu Ser Glu Ser Phe Val Leu Asp Phe Ser Gln Pro SerAla 105 110 115 gat tac tta gac ttt aga aat cga ctt cag gcc gac cac gtctgc ctt 437 Asp Tyr Leu Asp Phe Arg Asn Arg Leu Gln Ala Asp His Val CysLeu 120 125 130 135 gag aac tgt gtg ctc aag gac aag gcc att gca ggc actgtg aag gtt 485 Glu Asn Cys Val Leu Lys Asp Lys Ala Ile Ala Gly Thr ValLys Val 140 145 150 cag aac ctc gca ttt gag aag acc gtg aaa ata agg atgacg ttc gac 533 Gln Asn Leu Ala Phe Glu Lys Thr Val Lys Ile Arg Met ThrPhe Asp 155 160 165 acc tgg aag agc tac aca gac ttt cct tgt cag tac gtgaag gac act 581 Thr Trp Lys Ser Tyr Thr Asp Phe Pro Cys Gln Tyr Val LysAsp Thr 170 175 180 tat gcc ggt tca gac agg gac acg ttc tcc ttc gac atcagc ttg ccc 629 Tyr Ala Gly Ser Asp Arg Asp Thr Phe Ser Phe Asp Ile SerLeu Pro 185 190 195 gag aag att cag tct tat gaa aga atg gag ttt gct gtgtac tac gag 677 Glu Lys Ile Gln Ser Tyr Glu Arg Met Glu Phe Ala Val TyrTyr Glu 200 205 210 215 tgc aat gga cag acg tac tgg gac agc aac aga ggcaag aac tat agg 725 Cys Asn Gly Gln Thr Tyr Trp Asp Ser Asn Arg Gly LysAsn Tyr Arg 220 225 230 atc atc cgg gct gag tta aaa tct acc cag gga atgacc aag ccc cac 773 Ile Ile Arg Ala Glu Leu Lys Ser Thr Gln Gly Met ThrLys Pro His 235 240 245 agt gga ccg gat ttg gga ata tcc ttt gac cag ttcgga agc cct cgg 821 Ser Gly Pro Asp Leu Gly Ile Ser Phe Asp Gln Phe GlySer Pro Arg 250 255 260 tgt tcc tat ggt ctg ttt cca gag tgg cca agt tactta gga tat gaa 869 Cys Ser Tyr Gly Leu Phe Pro Glu Trp Pro Ser Tyr LeuGly Tyr Glu 265 270 275 aag cta ggg ccc tac tac tagtgactgc aggtgacagggcgtggcgga 917 Lys Leu Gly Pro Tyr Tyr 280 285 gctgccaca 926 12 285 PRTHomo sapiens 12 Met Met Ala Val Asp Ile Glu Tyr Arg Tyr Asn Cys Met AlaPro Ser 1 5 10 15 Leu Arg Gln Glu Arg Phe Ala Phe Lys Ile Ser Pro LysPro Ser Lys 20 25 30 Pro Leu Arg Pro Cys Ile Gln Leu Ser Ser Lys Asn GluAla Ser Gly 35 40 45 Met Val Ala Pro Ala Val Gln Glu Lys Lys Val Lys LysArg Val Ser 50 55 60 Phe Ala Asp Asn Gln Gly Leu Ala Leu Thr Met Val LysVal Phe Ser 65 70 75 80 Glu Phe Asp Asp Pro Leu Asp Met Pro Phe Asn IleThr Glu Leu Leu 85 90 95 Asp Asn Ile Val Ser Leu Thr Thr Ala Glu Ser GluSer Phe Val Leu 100 105 110 Asp Phe Ser Gln Pro Ser Ala Asp Tyr Leu AspPhe Arg Asn Arg Leu 115 120 125 Gln Ala Asp His Val Cys Leu Glu Asn CysVal Leu Lys Asp Lys Ala 130 135 140 Ile Ala Gly Thr Val Lys Val Gln AsnLeu Ala Phe Glu Lys Thr Val 145 150 155 160 Lys Ile Arg Met Thr Phe AspThr Trp Lys Ser Tyr Thr Asp Phe Pro 165 170 175 Cys Gln Tyr Val Lys AspThr Tyr Ala Gly Ser Asp Arg Asp Thr Phe 180 185 190 Ser Phe Asp Ile SerLeu Pro Glu Lys Ile Gln Ser Tyr Glu Arg Met 195 200 205 Glu Phe Ala ValTyr Tyr Glu Cys Asn Gly Gln Thr Tyr Trp Asp Ser 210 215 220 Asn Arg GlyLys Asn Tyr Arg Ile Ile Arg Ala Glu Leu Lys Ser Thr 225 230 235 240 GlnGly Met Thr Lys Pro His Ser Gly Pro Asp Leu Gly Ile Ser Phe 245 250 255Asp Gln Phe Gly Ser Pro Arg Cys Ser Tyr Gly Leu Phe Pro Glu Trp 260 265270 Pro Ser Tyr Leu Gly Tyr Glu Lys Leu Gly Pro Tyr Tyr 275 280 285 13551 DNA Homo sapiens CDS (38)..(505) 13 ctgtctcctg cattctcctg aaaccttcatccacaca atg cct ccc aac ctc act 55 Met Pro Pro Asn Leu Thr 1 5 ggc tactac cgc ttt gtc tcg cag aag aac atg gag gac tac ctg caa 103 Gly Tyr TyrArg Phe Val Ser Gln Lys Asn Met Glu Asp Tyr Leu Gln 10 15 20 gcc cta aacatc agc ttg gct gtg cgg aag atc gcg ctg ctg ctg aag 151 Ala Leu Asn IleSer Leu Ala Val Arg Lys Ile Ala Leu Leu Leu Lys 25 30 35 ccg gac aag gagatc gaa cac cag ggc aac cac atg acg gtg agg acg 199 Pro Asp Lys Glu IleGlu His Gln Gly Asn His Met Thr Val Arg Thr 40 45 50 ctc agc acc ttc cgaaac tac act gtg cag ttt gat gtg gga gtg gag 247 Leu Ser Thr Phe Arg AsnTyr Thr Val Gln Phe Asp Val Gly Val Glu 55 60 65 70 ttt gag gag gac ctcagg agc gtg gac gga cga aaa tgc cag atc tca 295 Phe Glu Glu Asp Leu ArgSer Val Asp Gly Arg Lys Cys Gln Ile Ser 75 80 85 ttc gtc ggt tcg gat ccaagc cag ttc tgt ggt cag caa ggc tcc cct 343 Phe Val Gly Ser Asp Pro SerGln Phe Cys Gly Gln Gln Gly Ser Pro 90 95 100 ctg ggc agg ccc cct ggtcag agg gag ttt gta tcc tca ggg agg agt 391 Leu Gly Arg Pro Pro Gly GlnArg Glu Phe Val Ser Ser Gly Arg Ser 105 110 115 ttg cgg ctg acc ttc cgcaca cag cct tcc tcg gag aac aag act gcc 439 Leu Arg Leu Thr Phe Arg ThrGln Pro Ser Ser Glu Asn Lys Thr Ala 120 125 130 cac ctc cac aag ggc ttcctg gcc ctc tac caa acc gtg gcc tta agt 487 His Leu His Lys Gly Phe LeuAla Leu Tyr Gln Thr Val Ala Leu Ser 135 140 145 150 gga agc ttg agt gacagc tgaggctggg gactcaggga cacctgggct 535 Gly Ser Leu Ser Asp Ser 155ggatcccagc cctgcc 551 14 156 PRT Homo sapiens 14 Met Pro Pro Asn Leu ThrGly Tyr Tyr Arg Phe Val Ser Gln Lys Asn 1 5 10 15 Met Glu Asp Tyr LeuGln Ala Leu Asn Ile Ser Leu Ala Val Arg Lys 20 25 30 Ile Ala Leu Leu LeuLys Pro Asp Lys Glu Ile Glu His Gln Gly Asn 35 40 45 His Met Thr Val ArgThr Leu Ser Thr Phe Arg Asn Tyr Thr Val Gln 50 55 60 Phe Asp Val Gly ValGlu Phe Glu Glu Asp Leu Arg Ser Val Asp Gly 65 70 75 80 Arg Lys Cys GlnIle Ser Phe Val Gly Ser Asp Pro Ser Gln Phe Cys 85 90 95 Gly Gln Gln GlySer Pro Leu Gly Arg Pro Pro Gly Gln Arg Glu Phe 100 105 110 Val Ser SerGly Arg Ser Leu Arg Leu Thr Phe Arg Thr Gln Pro Ser 115 120 125 Ser GluAsn Lys Thr Ala His Leu His Lys Gly Phe Leu Ala Leu Tyr 130 135 140 GlnThr Val Ala Leu Ser Gly Ser Leu Ser Asp Ser 145 150 155 15 817 DNA Homosapiens CDS (38)..(442) 15 ctgtctcctg cattctcctg aaaccttcat ccacaca atgcct ccc aac ctc act 55 Met Pro Pro Asn Leu Thr 1 5 ggc tac tac cgc tttgtc tcg cag aag aac atg gag gac tac ctg caa 103 Gly Tyr Tyr Arg Phe ValSer Gln Lys Asn Met Glu Asp Tyr Leu Gln 10 15 20 gcc cta aac atc agc ttggct gtg cgg aag atc gcg ctg ctg ctg aag 151 Ala Leu Asn Ile Ser Leu AlaVal Arg Lys Ile Ala Leu Leu Leu Lys 25 30 35 ccg gac aag gag atc gaa caccag ggc aac cac atg acg gtg agg acg 199 Pro Asp Lys Glu Ile Glu His GlnGly Asn His Met Thr Val Arg Thr 40 45 50 ctc agc acc ttc cga aac tac actgtg cag ttt gat gtg gga gtg gag 247 Leu Ser Thr Phe Arg Asn Tyr Thr ValGln Phe Asp Val Gly Val Glu 55 60 65 70 ttt gag gag gac ctc agg agc gtggac gga cga aaa tgc cag acc ata 295 Phe Glu Glu Asp Leu Arg Ser Val AspGly Arg Lys Cys Gln Thr Ile 75 80 85 gta acc tgg gag gag gag cac ctg gtgtgt gtg cag aaa ggg gag gtc 343 Val Thr Trp Glu Glu Glu His Leu Val CysVal Gln Lys Gly Glu Val 90 95 100 ccc aac cgg ggc tgg aga cac tgg ctggag gga gag ttg ctg tat ctg 391 Pro Asn Arg Gly Trp Arg His Trp Leu GluGly Glu Leu Leu Tyr Leu 105 110 115 gaa ctg act gca agg gat gca gtg tgcgag cag gtc ttc agg aag gtc 439 Glu Leu Thr Ala Arg Asp Ala Val Cys GluGln Val Phe Arg Lys Val 120 125 130 aga tagccggaga ggagccaaga tccctccagacagcaccagc tcacagacgc 492 Arg 135 tcttgttgtg cccccttcaa gcccagattgtgccagatct cattcgtcgg ttcggatcca 552 agccagttct gtggtcagca aggctcccctctgggcaggc cccctggtca gagggagttt 612 gtatcctcag ggaggagttt gcggctgaccttccgcacac agccttcctc ggagaacaag 672 actgcccacc tccacaaggg cttcctggccctctaccaaa ccgtgggtga gtgtccctcc 732 tgggggtgca gggagggagc ctctgttcccagccatgacc ctggtatctt caagccttaa 792 gtggaagctt gagtgacagc tgagg 817 16135 PRT Homo sapiens 16 Met Pro Pro Asn Leu Thr Gly Tyr Tyr Arg Phe ValSer Gln Lys Asn 1 5 10 15 Met Glu Asp Tyr Leu Gln Ala Leu Asn Ile SerLeu Ala Val Arg Lys 20 25 30 Ile Ala Leu Leu Leu Lys Pro Asp Lys Glu IleGlu His Gln Gly Asn 35 40 45 His Met Thr Val Arg Thr Leu Ser Thr Phe ArgAsn Tyr Thr Val Gln 50 55 60 Phe Asp Val Gly Val Glu Phe Glu Glu Asp LeuArg Ser Val Asp Gly 65 70 75 80 Arg Lys Cys Gln Thr Ile Val Thr Trp GluGlu Glu His Leu Val Cys 85 90 95 Val Gln Lys Gly Glu Val Pro Asn Arg GlyTrp Arg His Trp Leu Glu 100 105 110 Gly Glu Leu Leu Tyr Leu Glu Leu ThrAla Arg Asp Ala Val Cys Glu 115 120 125 Gln Val Phe Arg Lys Val Arg 130135 17 21 DNA Artificial Sequence Ag765 Forward Primer 17 ccaacgtgaagggagctata t 21 18 26 DNA Artificial Sequence Ag765 Probe Primer 18tgctgacacc actacacatg tcacaa 26 19 21 DNA Artificial Sequence Ag765Reverse Primer 19 ccagccccta aaattctcat c 21 20 22 DNA ArtificialSequence Ag1387 Forward Primer 20 ctgaaacctt catccacaca at 22 21 26 DNAArtificial Sequence Ag1387 Probe Primer 21 tcactggcta ctaccgcttt gtctcg26 22 22 DNA Artificial Sequence Ag1387 Reverse Primer 22 gcaggtagtcctccatgttc tt 22 23 218 DNA Homo sapiens 23 ctgaagtcac aggccctgcctctggctttt gcaggagaat tactacaagc tcctagccca 60 ggacacctgt ctgccctgcgactgcttccc ccatggctcc cacagccgca cttgcgacat 120 ggccaccggg cagtgtgcctgcaagcccgg cgtcatcggc cgccagtgca accgctgcga 180 caacccgttt gccgaggtcaccacgctcgg ctgtgaag 218 24 220 DNA Artificial Sequence ConsensusSequence 24 ctgnantnan annnncngcc nntgncnntn gcanggagaa ttactacaagctcctagccc 60 aggacacctg tctgccctgc gactgcttcc cccatggctc ccacagccgcacttgcgaca 120 tggccaccgg gcagtgtgcc tgcaagcccg gcgtcatcgg ccgccagtgcaaccgctgcg 180 nacaacccgt ttgccgaggt caccacgctc ggctgtgaag 220 25 3034PRT Mus musculus 25 Met Ala Pro Ser Ser Pro Arg Val Leu Pro Ala Leu ValLeu Leu Ala 1 5 10 15 Ala Ala Ala Leu Pro Ala Leu Glu Leu Gly Ala AlaAla Trp Glu Leu 20 25 30 Arg Val Pro Gly Gly Ala Arg Ala Phe Ala Leu GlyPro Gly Trp Ser 35 40 45 Tyr Arg Leu Asp Thr Thr Arg Thr Pro Arg Glu LeuLeu Asp Val Ser 50 55 60 Arg Glu Gly Pro Ala Ala Gly Arg Arg Leu Gly LeuGly Ala Gly Thr 65 70 75 80 Leu Gly Cys Ala Arg Leu Ala Gly Arg Leu LeuPro Leu Gln Val Arg 85 90 95 Leu Val Ala Arg Gly Ala Pro Thr Ala Pro SerLeu Val Leu Arg Ala 100 105 110 Arg Ala Tyr Gly Ala Arg Cys Gly Val ArgLeu Leu Arg Arg Ser Ala 115 120 125 Arg Gly Ala Glu Leu Arg Ser Pro AlaVal Arg Ser Val Pro Gly Leu 130 135 140 Gly Asp Ala Leu Cys Phe Pro AlaAla Gly Gly Gly Ala Ala Ser Leu 145 150 155 160 Thr Ser Val Leu Glu AlaIle Thr Asn Phe Pro Ala Cys Ser Cys Pro 165 170 175 Pro Val Ala Gly ThrGly Cys Arg Arg Gly Pro Ile Cys Leu Arg Pro 180 185 190 Gly Gly Ser AlaGlu Leu Arg Leu Val Cys Ala Leu Gly Arg Ala Ala 195 200 205 Gly Ala ValTrp Val Glu Leu Val Ile Gln Ala Thr Ser Gly Thr Pro 210 215 220 Ser GluSer Pro Ser Val Ser Pro Ser Leu Leu Asn Leu Ser Gln Pro 225 230 235 240Arg Ala Gly Val Val Arg Arg Ser Arg Arg Gly Thr Gly Ser Ser Thr 245 250255 Ser Pro Gln Phe Pro Leu Pro Ser Tyr Gln Val Ser Val Pro Glu Asn 260265 270 Glu Pro Ala Gly Thr Ala Val Ile Glu Leu Arg Ala His Asp Pro Asp275 280 285 Glu Gly Asp Ala Gly Arg Leu Ser Tyr Gln Met Glu Ala Leu PheAsp 290 295 300 Glu Arg Ser Asn Gly Tyr Phe Leu Ile Asp Ala Ala Thr GlyAla Val 305 310 315 320 Thr Thr Ala Arg Ser Leu Asp Arg Glu Thr Lys AspThr His Val Leu 325 330 335 Lys Val Ser Ala Val Asp His Gly Ser Pro ArgArg Ser Ala Ala Thr 340 345 350 Tyr Leu Thr Val Thr Val Ser Asp Thr AsnAsp His Ser Pro Val Phe 355 360 365 Glu Gln Ser Glu Tyr Arg Glu Arg IleArg Glu Asn Leu Glu Val Gly 370 375 380 Tyr Glu Val Leu Thr Ile Arg AlaThr Asp Gly Asp Ala Pro Ser Asn 385 390 395 400 Ala Asn Met Arg Tyr ArgLeu Leu Glu Gly Ala Gly Gly Val Phe Glu 405 410 415 Ile Asp Ala Arg SerGly Val Val Arg Thr Arg Ala Val Val Asp Arg 420 425 430 Glu Glu Ala AlaGlu Tyr Gln Leu Leu Val Glu Ala Asn Asp Gln Gly 435 440 445 Arg Asn ProGly Pro Leu Ser Ala Ser Ala Thr Val His Ile Val Val 450 455 460 Glu AspGlu Asn Asp Asn Tyr Pro Gln Phe Ser Glu Lys Arg Tyr Val 465 470 475 480Val Gln Val Pro Glu Asp Val Ala Val Asn Thr Ala Val Leu Arg Val 485 490495 Gln Ala Thr Asp Arg Asp Gln Gly Gln Asn Ala Ala Ile His Tyr Ser 500505 510 Ile Val Ser Gly Asn Leu Lys Gly Gln Phe Tyr Leu His Ser Leu Ser515 520 525 Gly Ser Leu Asp Val Ile Asn Pro Leu Asp Phe Glu Ala Ile ArgGlu 530 535 540 Tyr Thr Leu Arg Ile Lys Ala Gln Asp Gly Gly Arg Pro ProLeu Ile 545 550 555 560 Asn Ser Ser Gly Leu Val Ser Val Gln Val Leu AspVal Asn Asp Asn 565 570 575 Ala Pro Ile Phe Val Ser Ser Pro Phe Gln AlaAla Val Leu Glu Asn 580 585 590 Val Pro Leu Gly His Ser Val Leu His IleGln Ala Val Asp Ala Asp 595 600 605 Ala Gly Glu Asn Ala Arg Leu Gln TyrArg Leu Val Asp Thr Ala Ser 610 615 620 Thr Ile Val Gly Gly Ser Ser ValAsp Ser Glu Asn Pro Ala Ser Ala 625 630 635 640 Pro Asp Phe Pro Phe GlnIle His Asn Ser Ser Gly Trp Ile Thr Val 645 650 655 Cys Ala Glu Leu AspArg Glu Glu Val Glu His Tyr Ser Phe Gly Val 660 665 670 Glu Ala Val AspHis Gly Ser Pro Ala Met Ser Ser Ser Ala Ser Val 675 680 685 Ser Ile ThrVal Leu Asp Val Asn Asp Asn Asp Pro Met Phe Thr Gln 690 695 700 Pro ValTyr Glu Leu Arg Leu Asn Glu Asp Ala Ala Val Gly Ser Ser 705 710 715 720Val Leu Thr Leu Arg Ala Arg Asp Arg Asp Ala Asn Ser Val Ile Thr 725 730735 Tyr Gln Leu Thr Gly Gly Asn Thr Arg Asn Arg Phe Ala Leu Ser Ser 740745 750 Gln Ser Gly Gly Gly Leu Ile Thr Leu Ala Leu Pro Leu Asp Tyr Lys755 760 765 Gln Glu Arg Gln Tyr Val Leu Ala Val Thr Ala Ser Asp Gly ThrArg 770 775 780 Ser His Thr Ala Gln Val Phe Ile Asn Val Thr Asp Ala AsnThr His 785 790 795 800 Arg Pro Val Phe Gln Ser Ser His Tyr Thr Val SerVal Ser Glu Asp 805 810 815 Arg Pro Val Gly Thr Ser Ile Ala Thr Ile SerAla Thr Asp Glu Asp 820 825 830 Thr Gly Glu Asn Ala Arg Ile Thr Tyr ValLeu Glu Asp Pro Val Pro 835 840 845 Gln Phe Arg Ile Asp Pro Asp Thr GlyThr Ile Tyr Thr Met Thr Glu 850 855 860 Leu Asp Tyr Glu Asp Gln Ala AlaTyr Thr Leu Ala Ile Thr Ala Gln 865 870 875 880 Asp Asn Gly Ile Pro GlnLys Ser Asp Thr Thr Ser Leu Glu Ile Leu 885 890 895 Ile Leu Asp Ala AsnAsp Asn Ala Pro Arg Phe Leu Arg Asp Phe Tyr 900 905 910 Gln Gly Ser ValPhe Glu Asp Ala Pro Pro Ser Thr Ser Val Leu Gln 915 920 925 Val Ser AlaThr Asp Arg Asp Ser Gly Pro Asn Gly Arg Leu Leu Tyr 930 935 940 Thr PheGln Gly Gly Asp Asp Gly Asp Gly Asp Phe Tyr Ile Glu Pro 945 950 955 960Thr Ser Gly Val Ile Arg Thr Gln Arg Arg Leu Asp Arg Glu Asn Val 965 970975 Ala Val Tyr Asn Leu Trp Ala Leu Ala Val Asp Arg Gly Ser Pro Asn 980985 990 Pro Leu Ser Ala Ser Val Gly Ile Gln Val Ser Val Leu Asp Ile Asn995 1000 1005 Asp Asn Pro Pro Val Phe Glu Lys Asp Glu Leu Glu Leu PheVal Glu 1010 1015 1020 Glu Asn Ser Pro Val Gly Ser Val Val Ala Arg IleArg Ala Asn Asp 1025 1030 1035 1040 Pro Asp Glu Gly Pro Asn Ala Gln IleIle Tyr Gln Ile Val Glu Gly 1045 1050 1055 Asn Val Pro Glu Val Phe GlnLeu Asp Leu Leu Ser Gly Asp Leu Arg 1060 1065 1070 Ala Leu Val Glu LeuAsp Phe Glu Val Arg Arg Asp Tyr Met Leu Val 1075 1080 1085 Val Gln AlaThr Ser Ala Pro Leu Val Ser Arg Ala Thr Val His Ile 1090 1095 1100 ArgLeu Leu Asp Gln Asn Asp Asn Pro Pro Glu Leu Pro Asp Phe Gln 1105 11101115 1120 Ile Leu Phe Asn Asn Tyr Val Thr Asn Lys Ser Asn Ser Phe ProSer 1125 1130 1135 Gly Val Ile Gly Arg Ile Pro Ala His Asp Pro Asp LeuSer Asp Ser 1140 1145 1150 Leu Asn Tyr Thr Phe Leu Gln Gly Asn Glu LeuSer Leu Leu Leu Leu 1155 1160 1165 Asp Pro Ala Thr Gly Glu Leu Gln LeuSer Arg Asp Leu Asp Asn Asn 1170 1175 1180 Arg Pro Leu Glu Ala Leu MetGlu Val Ser Val Ser Asp Gly Ile His 1185 1190 1195 1200 Ser Val Thr AlaLeu Cys Thr Leu Arg Val Thr Ile Ile Thr Asp Asp 1205 1210 1215 Met LeuThr Asn Ser Ile Thr Val Arg Leu Glu Asn Met Ser Gln Glu 1220 1225 1230Lys Phe Leu Ser Pro Leu Leu Ser Leu Phe Val Glu Gly Val Ala Thr 12351240 1245 Val Leu Ser Thr Thr Lys Asp Asp Ile Phe Val Phe Asn Ile GlnAsn 1250 1255 1260 Asp Thr Asp Val Ser Ser Asn Ile Leu Asn Val Thr PheSer Ala Leu 1265 1270 1275 1280 Leu Pro Gly Gly Thr Arg Gly Arg Phe PhePro Ser Glu Asp Leu Gln 1285 1290 1295 Glu Gln Ile Tyr Leu Asn Arg ThrLeu Leu Thr Thr Ile Ser Ala Gln 1300 1305 1310 Arg Val Leu Pro Phe AspAsp Asn Ile Cys Leu Arg Glu Pro Cys Glu 1315 1320 1325 Asn Tyr Met LysCys Val Ser Val Leu Arg Phe Asp Ser Ser Ala Pro 1330 1335 1340 Phe IleSer Ser Thr Thr Val Leu Phe Arg Pro Ile His Pro Ile Thr 1345 1350 13551360 Gly Leu Arg Cys Arg Cys Pro Pro Gly Phe Thr Gly Asp Tyr Cys Glu1365 1370 1375 Thr Glu Ile Asp Leu Cys Tyr Ser Asn Pro Cys Gly Ala AsnGly Arg 1380 1385 1390 Cys Arg Ser Arg Glu Gly Gly Tyr Thr Cys Glu CysPhe Glu Asp Phe 1395 1400 1405 Thr Gly Glu His Cys Gln Val Asn Val ArgSer Gly Arg Cys Ala Ser 1410 1415 1420 Gly Val Cys Lys Asn Gly Gly ThrCys Val Asn Leu Leu Ile Gly Gly 1425 1430 1435 1440 Phe His Cys Val CysPro Pro Gly Glu Tyr Glu His Pro Tyr Cys Glu 1445 1450 1455 Val Ser ThrArg Ser Phe Pro Pro Gln Ser Phe Val Thr Phe Arg Gly 1460 1465 1470 LeuArg Gln Arg Phe His Phe Thr Val Ser Leu Ala Phe Ala Thr Gln 1475 14801485 Asp Arg Asn Ala Leu Leu Leu Tyr Asn Gly Arg Phe Asn Glu Lys His1490 1495 1500 Asp Phe Ile Ala Leu Glu Ile Val Glu Glu Gln Leu Gln LeuThr Phe 1505 1510 1515 1520 Ser Ala Gly Glu Thr Thr Thr Thr Val Thr ProGln Val Pro Gly Gly 1525 1530 1535 Val Ser Asp Gly Arg Trp His Ser ValLeu Val Gln Tyr Tyr Asn Lys 1540 1545 1550 Pro Asn Ile Gly His Leu GlyLeu Pro His Gly Pro Ser Gly Glu Lys 1555 1560 1565 Val Ala Val Val ThrVal Asp Asp Cys Asp Ala Ala Val Ala Val His 1570 1575 1580 Phe Gly SerTyr Val Gly Asn Tyr Ser Cys Ala Ala Gln Gly Thr Gln 1585 1590 1595 1600Ser Gly Ser Lys Lys Ser Leu Asp Leu Thr Gly Pro Leu Leu Leu Gly 16051610 1615 Gly Val Pro Asn Leu Pro Glu Asp Phe Pro Val His Ser Arg GlnPhe 1620 1625 1630 Val Gly Cys Met Arg Asn Leu Ser Ile Asp Gly Arg IleVal Asp Met 1635 1640 1645 Ala Ala Phe Ile Ala Asn Asn Gly Thr Arg AlaGly Cys Ala Ser Gln 1650 1655 1660 Arg Asn Phe Cys Asp Gly Thr Ser CysGln Asn Gly Gly Thr Cys Val 1665 1670 1675 1680 Asn Arg Trp Asn Thr TyrLeu Cys Glu Cys Pro Leu Arg Phe Gly Gly 1685 1690 1695 Lys Asn Cys GluGln Ala Met Pro His Pro Gln Arg Phe Thr Gly Glu 1700 1705 1710 Ser ValVal Leu Trp Ser Asp Leu Asp Ile Thr Ile Ser Val Pro Trp 1715 1720 1725Tyr Leu Gly Leu Met Phe Arg Thr Arg Lys Glu Asp Gly Val Leu Met 17301735 1740 Glu Ala Thr Ala Gly Thr Ser Ser Arg Leu His Leu Gln Ile LeuAsn 1745 1750 1755 1760 Ser Tyr Ile Arg Phe Glu Val Ser Tyr Gly Pro SerAsp Val Ala Ser 1765 1770 1775 Met Gln Leu Ser Lys Ser Arg Ile Thr AspGly Gly Trp His His Leu 1780 1785 1790 Leu Ile Glu Leu Arg Ser Ala LysGlu Gly Lys Asp Ile Lys Tyr Leu 1795 1800 1805 Ala Val Met Thr Leu AspTyr Gly Met Asp Gln Ser Thr Val Gln Ile 1810 1815 1820 Gly Asn Gln LeuPro Gly Leu Lys Met Arg Thr Ile Val Ile Gly Gly 1825 1830 1835 1840 ValThr Glu Asp Lys Val Ser Val Arg His Gly Phe Arg Gly Cys Met 1845 18501855 Gln Gly Val Arg Met Gly Glu Thr Ser Thr Asn Ile Ala Thr Leu Asn1860 1865 1870 Met Asn Asp Ala Leu Lys Val Arg Val Lys Asp Gly Cys AspVal Glu 1875 1880 1885 Asp Pro Cys Ala Ser Ser Pro Cys Pro Pro His ArgPro Cys Arg Asp 1890 1895 1900 Thr Trp Asp Ser Tyr Ser Cys Ile Cys AspArg Gly Tyr Phe Gly Lys 1905 1910 1915 1920 Lys Cys Val Asp Ala Cys LeuLeu Asn Pro Cys Lys His Val Ala Ala 1925 1930 1935 Cys Val Arg Ser ProAsn Thr Pro Arg Gly Tyr Ser Cys Glu Cys Gly 1940 1945 1950 Pro Gly HisTyr Gly Gln Tyr Cys Glu Asn Lys Val Asp Leu Pro Cys 1955 1960 1965 ProLys Gly Trp Trp Gly Asn Pro Val Cys Gly Pro Cys His Cys Ala 1970 19751980 Val Ser Gln Gly Phe Asp Pro Asp Cys Asn Lys Thr Asn Gly Gln Cys1985 1990 1995 2000 Gln Cys Lys Glu Asn Tyr Tyr Lys Pro Pro Ala Gln AspAla Cys Leu 2005 2010 2015 Pro Cys Asp Cys Phe Pro His Gly Ser His SerArg Ala Cys Asp Met 2020 2025 2030 Asp Thr Gly Gln Cys Ala Cys Lys ProGly Val Ile Gly Arg Gln Cys 2035 2040 2045 Asn Arg Cys Asp Asn Pro PheAla Glu Val Thr Ser Leu Gly Cys Glu 2050 2055 2060 Val Ile Tyr Asn GlyCys Pro Arg Ala Phe Glu Ala Gly Ile Trp Trp 2065 2070 2075 2080 Pro GlnThr Lys Phe Gly Gln Pro Ala Ala Val Pro Cys Pro Lys Gly 2085 2090 2095Ser Val Gly Asn Ala Val Arg His Cys Ser Gly Glu Lys Gly Trp Leu 21002105 2110 Pro Pro Glu Leu Phe Asn Cys Thr Ser Gly Ser Phe Val Asp LeuLys 2115 2120 2125 Ala Leu Asn Glu Lys Leu Asn Arg Asn Glu Thr Arg MetAsp Gly Asn 2130 2135 2140 Arg Ser Leu Arg Leu Ala Lys Ala Leu Arg AsnAla Thr Gln Gly Asn 2145 2150 2155 2160 Ser Thr Leu Phe Gly Asn Asp ValArg Thr Ala Tyr Gln Leu Leu Ala 2165 2170 2175 Arg Ile Leu Gln His GluSer Arg Gln Gln Gly Phe Asp Leu Ala Ala 2180 2185 2190 Thr Arg Glu AlaAsn Phe His Glu Asp Val Val His Thr Gly Ser Ala 2195 2200 2205 Leu LeuAla Pro Ala Thr Glu Ala Ser Trp Glu Gln Ile Gln Arg Ser 2210 2215 2220Glu Ala Gly Ala Ala Gln Leu Leu Arg His Phe Glu Ala Tyr Phe Ser 22252230 2235 2240 Asn Val Ala Arg Asn Val Lys Arg Thr Tyr Leu Arg Pro PheVal Ile 2245 2250 2255 Val Thr Ala Asn Met Ile Leu Ala Val Asp Ile PheAsp Lys Leu Asn 2260 2265 2270 Phe Thr Gly Ala Gln Val Pro Arg Phe GluAsp Ile Gln Glu Glu Leu 2275 2280 2285 Pro Arg Glu Leu Glu Ser Ser ValSer Phe Pro Ala Asp Thr Phe Lys 2290 2295 2300 Pro Pro Glu Lys Lys GluGly Pro Val Val Arg Leu Thr Asn Arg Arg 2305 2310 2315 2320 Thr Thr ProLeu Thr Ala Gln Pro Glu Pro Arg Ala Glu Arg Glu Thr 2325 2330 2335 SerSer Ser Arg Arg Arg Arg His Pro Asp Glu Pro Gly Gln Phe Ala 2340 23452350 Val Ala Leu Val Val Ile Tyr Arg Thr Leu Gly Gln Leu Leu Pro Glu2355 2360 2365 His Tyr Asp Pro Asp His Arg Ser Leu Arg Leu Pro Asn ArgPro Val 2370 2375 2380 Ile Asn Thr Pro Val Val Ser Ala Met Val Tyr SerGlu Gly Thr Pro 2385 2390 2395 2400 Leu Pro Ser Ser Leu Gln Arg Pro IleLeu Val Glu Phe Ser Leu Leu 2405 2410 2415 26 120 PRT ArtificialSequence Consensus Sequence 26 Tyr Xaa Gly Xaa Xaa Cys Val Asp Ala CysXaa Leu Asn Pro Cys Xaa 1 5 10 15 Xaa Xaa Xaa Ala Cys Val Arg Ser ProXaa Xaa Pro Xaa Gly Tyr Xaa 20 25 30 Cys Glu Cys Gly Pro Xaa His Tyr GlyXaa Tyr Cys Glu Asn Lys Xaa 35 40 45 Asp Leu Pro Cys Pro Xaa Gly Trp TrpGly Asn Pro Val Cys Gly Pro 50 55 60 Cys His Cys Ala Val Ser Xaa Gly PheAsp Pro Asp Cys Asn Lys Thr 65 70 75 80 Asn Gly Gln Cys Gln Cys Lys GluAsn Tyr Tyr Lys Xaa Xaa Ala Gln 85 90 95 Asp Xaa Cys Leu Pro Cys Asp CysPhe Pro His Gly Ser His Ser Arg 100 105 110 Xaa Cys Asp Met Xaa Thr GlyGln 115 120 27 1713 PRT Artificial Sequence Consensus Sequence 27 MetAla Pro Xaa Xaa Pro Xaa Val Leu Pro Xaa Leu Xaa Leu Ala Ala 1 5 10 15Ala Ala Xaa Leu Pro Ala Xaa Xaa Leu Xaa Ala Ala Ala Trp Glu Xaa 20 25 30Arg Val Pro Gly Gly Xaa Arg Ala Phe Ala Leu Xaa Pro Gly Xaa Xaa 35 40 45Tyr Xaa Xaa Xaa Xaa Xaa Thr Pro Arg Xaa Arg Glu Xaa Xaa Xaa Xaa 50 55 60Gly Arg Xaa Xaa Gly Ala Gly Xaa Xaa Xaa Xaa Xaa Xaa Xaa Ala Gly 65 70 7580 Arg Xaa Leu Pro Leu Gln Val Arg Leu Val Ala Arg Xaa Ala Pro Thr 85 9095 Ala Xaa Ser Xaa Xaa Leu Arg Ala Arg Xaa Xaa Xaa Xaa Xaa Cys Xaa 100105 110 Gly Xaa Arg Xaa Xaa Arg Xaa Xaa Xaa Xaa Gly Ala Xaa Leu Xaa Xaa115 120 125 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Gly Ala Leu Cys Phe Pro XaaXaa 130 135 140 Gly Gly Xaa Ala Ala Xaa Xaa Xaa Ser Xaa Leu Xaa Ala XaaThr Xaa 145 150 155 160 Xaa Pro Ala Cys Xaa Cys Pro Pro Xaa Cys Xaa XaaXaa Pro Ile Cys 165 170 175 Leu Xaa Pro Gly Gly Ser Xaa Xaa Leu Arg LeuXaa Cys Ala Leu Xaa 180 185 190 Arg Ala Ala Gly Ala Val Xaa Val Xaa LeuXaa Xaa Xaa Ala Xaa Thr 195 200 205 Xaa Gly Thr Pro Ser Xaa Ser Pro SerXaa Ser Pro Xaa Leu Xaa Xaa 210 215 220 Asn Leu Xaa Xaa Xaa Arg Ala GlyXaa Xaa Arg Arg Xaa Arg Arg Gly 225 230 235 240 Thr Xaa Xaa Xaa Xaa SerXaa Xaa Phe Pro Xaa Pro Xaa Tyr Gln Val 245 250 255 Xaa Xaa Xaa Glu AsnGlu Pro Ala Gly Thr Xaa Xaa Xaa Xaa Leu Xaa 260 265 270 Ala His Xaa GluGly Xaa Xaa Xaa Arg Xaa Ser Tyr Xaa Met Glu Xaa 275 280 285 Leu Phe AspGlu Arg Ser Xaa Gly Tyr Phe Xaa Ile Asp Xaa Ala Thr 290 295 300 Gly AlaVal Xaa Thr Xaa Xaa Xaa Leu Asp Arg Glu Thr Lys Xaa Thr 305 310 315 320His Val Leu Xaa Val Xaa Ala Val Asp Xaa Xaa Xaa Pro Xaa Arg Ser 325 330335 Ala Xaa Thr Tyr Xaa Thr Val Val Asp Thr Asn Asp His Ser Pro Val 340345 350 Phe Glu Gln Ser Glu Tyr Arg Glu Arg Xaa Arg Glu Asn Leu Glu Val355 360 365 Gly Tyr Glu Val Leu Thr Ile Arg Ala Xaa Asp Xaa Asp Xaa ProXaa 370 375 380 Asn Ala Asn Xaa Arg Tyr Arg Xaa Leu Xaa Gly Ala Xaa XaaVal Phe 385 390 395 400 Xaa Xaa Xaa Xaa Xaa Ser Gly Val Val Thr Arg AlaVal Xaa Asp Arg 405 410 415 Glu Glu Ala Ala Glu Tyr Gln Leu Leu Val GluAla Asn Asp Gln Gly 420 425 430 Arg Asn Pro Gly Pro Leu Ser Ala Ala ThrVal Xaa Ile Xaa Val Glu 435 440 445 Asp Glu Asn Asp Asn Tyr Pro Gln PheSer Glu Xaa Tyr Val Val Gln 450 455 460 Val Pro Glu Asp Val Xaa Xaa AsnThr Ala Val Leu Arg Val Gln Ala 465 470 475 480 Thr Asp Arg Asp Gln GlyGln Asn Ala Ala Ile His Tyr Ser Ile Xaa 485 490 495 Ser Gly Asn Xaa XaaGly Gln Phe Tyr Leu His Ser Leu Ser Gly Xaa 500 505 510 Leu Asp Val IleAsn Pro Leu Xaa Asp Phe Glu Xaa Xaa Xaa Xaa Tyr 515 520 525 Xaa Leu IleLys Ala Gln Asp Gly Gly Arg Pro Pro Leu Ile Asn Ser 530 535 540 Ser GlyVal Ser Val Gln Val Leu Asp Val Asn Asp Asn Xaa Pro Ile 545 550 555 560Phe Val Ser Ser Pro Phe Gln Ala Val Leu Glu Asn Val Pro Leu Gly 565 570575 Xaa Xaa Val Xaa His Ile Gln Ala Val Asp Ala Asp Xaa Gly Glu Asn 580585 590 Ala Arg Leu Xaa Tyr Arg Leu Val Asp Thr Ala Ser Thr Xaa Xaa Gly595 600 605 Gly Xaa Ser Xaa Xaa Xaa Xaa Asn Pro Ala Xaa Pro Asp Phe ProPhe 610 615 620 Gln Ile His Asn Ser Ser Gly Trp Ile Thr Val Cys Ala GluLeu Asp 625 630 635 640 Arg Glu Glu Val Glu His Tyr Ser Phe Gly Val GluAla Val Asp His 645 650 655 Gly Ser Pro Xaa Met Ser Ser Ser Ser Val SerIle Thr Val Leu Asp 660 665 670 Val Asn Asp Asn Asp Pro Xaa Phe Thr GlnPro Tyr Glu Leu Arg Leu 675 680 685 Asn Glu Asp Ala Ala Val Xaa Gly SerSer Val Leu Thr Leu Xaa Ala 690 695 700 Arg Asp Arg Asp Ala Asn Ser ValIle Thr Tyr Gln Leu Thr Gly Gly 705 710 715 720 Asn Thr Arg Asn Arg PheAla Leu Ser Ser Gln Gly Gly Gly Leu Ile 725 730 735 Thr Leu Ala Leu ProLeu Asp Tyr Lys Gln Glu Xaa Gln Tyr Val Leu 740 745 750 Ala Val Xaa ThrAla Ser Asp Gly Thr Arg Ser His Thr Ala Xaa Val 755 760 765 Ile Asn ValThr Asp Ala Asn Thr His Arg Pro Val Phe Gln Ser Ser 770 775 780 His TyrThr Val Ser Val Ser Glu Asp Arg Pro Val Gly Thr Ser Ile 785 790 795 800Ala Thr Ser Ala Xaa Asp Glu Asp Thr Gly Glu Asn Ala Xaa Arg Ile 805 810815 Thr Tyr Val Xaa Xaa Asp Pro Val Pro Gln Phe Arg Ile Asp Pro Asp 820825 830 Xaa Gly Thr Xaa Tyr Thr Met Xaa Glu Leu Asp Tyr Glu Xaa Gln Xaa835 840 845 Ala Tyr Thr Leu Xaa Ile Xaa Ala Gln Asp Asn Gly Ile Pro GlnLys 850 855 860 Ser Asp Thr Thr Xaa Leu Glu Ile Leu Ile Leu Asp Ala AsnAsp Asn 865 870 875 880 Ala Pro Xaa Phe Leu Xaa Asp Phe Tyr Gln Gly SerXaa Phe Glu Asp 885 890 895 Ala Pro Pro Ser Thr Ser Xaa Leu Gln Val SerAla Thr Asp Arg Asp 900 905 910 Ser Gly Pro Asn Gly Arg Leu Leu Tyr ThrPhe Gln Gly Gly Asp Asp 915 920 925 Gly Asp Gly Asp Phe Tyr Ile Glu ProThr Ser Gly Val Ile Arg Thr 930 935 940 Gln Arg Arg Leu Asp Arg Glu AsnVal Ala Val Tyr Asn Leu Trp Ala 945 950 955 960 Leu Ala Val Asp Arg GlySer Pro Xaa Pro Leu Ser Ala Ser Val Xaa 965 970 975 Ile Gln Val Xaa XaaLeu Asp Ile Asn Asp Asn Xaa Pro Xaa Phe Glu 980 985 990 Lys Asp Glu LeuGlu Leu Phe Val Glu Glu Asn Xaa Pro Val Gly Ser 995 1000 1005 Val ValAla Xaa Ile Arg Ala Asn Asp Pro Asp Glu Gly Pro Asn Ala 1010 1015 1020Gln Ile Xaa Tyr Gln Ile Val Glu Gly Xaa Xaa Xaa Xaa Xaa Phe Gln 10251030 1035 1040 Leu Asp Leu Leu Xaa Gly Asp Leu Arg Ala Xaa Val Glu LeuAsp Phe 1045 1050 1055 Glu Val Arg Arg Xaa Tyr Xaa Leu Val Val Gln AlaThr Ser Ala Pro 1060 1065 1070 Leu Val Ser Arg Ala Thr Val His Ile XaaLeu Xaa Asp Gln Asn Asp 1075 1080 1085 Asn Pro Pro Xaa Leu Pro Asp PheGln Ile Leu Phe Asn Asn Tyr Val 1090 1095 1100 Thr Asn Lys Ser Asn SerPhe Pro Xaa Gly Xaa Val Ile Gly Ile Pro 1105 1110 1115 1120 Ala His AspPro Asp Xaa Ser Asp Ser Leu Asn Tyr Thr Phe Xaa Gln 1125 1130 1135 GlyAsn Glu Leu Xaa Leu Leu Leu Leu Asp Pro Ala Thr Gly Glu Leu 1140 11451150 Gln Leu Ser Arg Asp Leu Asp Asn Asn Arg Pro Leu Glu Ala Leu Met1155 1160 1165 Glu Val Ser Val Ser Asp Xaa Gly Ile His Ser Val Thr AlaCys Thr 1170 1175 1180 Leu Arg Val Thr Ile Ile Thr Asp Asp Met Leu ThrAsn Ser Ile Thr 1185 1190 1195 1200 Val Arg Leu Glu Asn Met Ser Gln GluLys Phe Leu Ser Pro Leu Leu 1205 1210 1215 Xaa Leu Phe Val Glu Gly ValAla Xaa Val Leu Ser Thr Thr Lys Asp 1220 1225 1230 Asp Xaa Phe Val PheAsn Xaa Gln Asn Asp Thr Asp Val Ser Ser Asn 1235 1240 1245 Ile Leu AsnVal Thr Phe Ser Ala Leu Leu Pro Gly Gly Xaa Arg Gly 1250 1255 1260 XaaPhe Phe Pro Ser Glu Asp Leu Gln Glu Gln Ile Tyr Leu Asn Arg 1265 12701275 1280 Thr Leu Leu Thr Thr Ile Ser Xaa Gln Arg Val Leu Pro Phe AspAsp 1285 1290 1295 Asn Ile Cys Leu Arg Glu Pro Cys Glu Asn Tyr Met LysCys Val Ser 1300 1305 1310 Val Leu Arg Phe Asp Ser Ser Ala Pro Phe XaaSer Ser Thr Thr Val 1315 1320 1325 Leu Phe Arg Pro Ile His Pro Ile XaaGly Leu Arg Cys Arg Cys Pro 1330 1335 1340 Pro Gly Phe Thr Gly Asp TyrCys Glu Thr Glu Ile Asp Leu Cys Tyr 1345 1350 1355 1360 Ser Pro Cys GlyAla Asn Gly Arg Cys Arg Ser Arg Glu Gly Gly Tyr 1365 1370 1375 Thr CysGlu Cys Phe Glu Asp Phe Thr Gly Glu His Cys Xaa Val Xaa 1380 1385 1390Xaa Arg Ser Gly Arg Cys Ala Xaa Gly Val Cys Lys Asn Gly Gly Thr 13951400 1405 Cys Val Asn Leu Leu Ile Gly Gly Phe His Cys Val Cys Pro ProGly 1410 1415 1420 Glu Tyr Glu Xaa Pro Tyr Cys Glu Val Xaa Thr Arg SerPhe Pro Pro 1425 1430 1435 1440 Gln Ser Phe Val Thr Phe Arg Gly Leu ArgGln Arg Phe His Phe Thr 1445 1450 1455 Xaa Ser Leu Xaa Phe Ala Thr GlnXaa Arg Asn Xaa Leu Leu Leu Tyr 1460 1465 1470 Asn Gly Arg Phe Asn GluLys His Asp Phe Ile Ala Leu Glu Ile Val 1475 1480 1485 Xaa Glu Gln XaaGln Leu Thr Phe Ser Ala Gly Glu Thr Thr Thr Thr 1490 1495 1500 Val XaaPro Xaa Val Pro Xaa Gly Val Ser Asp Gly Arg Trp His Ser 1505 1510 15151520 Val Xaa Val Gln Tyr Tyr Asn Lys Pro Asn Ile Gly His Leu Gly Leu1525 1530 1535 Pro His Gly Pro Ser Gly Glu Lys Xaa Ala Val Val Thr ValAsp Asp 1540 1545 1550 Cys Asp Xaa Xaa Xaa Ala Val Xaa Phe Gly Xaa XaaXaa Gly Asn Tyr 1555 1560 1565 Ser Cys Ala Ala Gln Gly Thr Gln Xaa GlySer Lys Lys Ser Leu Asp 1570 1575 1580 Leu Thr Gly Pro Leu Leu Leu GlyGly Val Pro Asn Leu Pro Glu Asp 1585 1590 1595 1600 Phe Pro Val His XaaArg Gln Phe Val Gly Cys Met Arg Asn Leu Ser 1605 1610 1615 Xaa Asp GlyXaa Xaa Val Asp Met Ala Xaa Phe Ile Ala Asn Asn Gly 1620 1625 1630 ThrArg Xaa Gly Cys Ala Xaa Xaa Arg Asn Phe Cys Asp Gly Xaa Xaa 1635 16401645 Cys Gln Asn Gly Gly Xaa Thr Cys Val Asn Arg Trp Asn Tyr Leu Cys1650 1655 1660 Glu Cys Pro Leu Arg Phe Gly Gly Lys Asn Cys Glu Gln AlaMet Pro 1665 1670 1675 1680 His Pro Gln Xaa Phe Xaa Gly Glu Ser Val ValXaa Trp Ser Asp Leu 1685 1690 1695 Xaa Ile Xaa Ile Ser Val Pro Trp TyrLeu Gly Leu Met Phe Arg Thr 1700 1705 1710 Arg 28 458 PRT ArtificialSequence Consensus Sequence 28 Val Arg Asn Ile Asp Thr Gly Pro Met ArgPhe Tyr Xaa Val Val Gly 1 5 10 15 Trp Gly Ile Pro Ala Ile Val Thr GlyLeu Ala Val Gly Leu Asp Pro 20 25 30 Gln Gly Tyr Gly Asn Pro Asp Phe CysTrp Leu Ser Leu Gln Asp Thr 35 40 45 Leu Ile Trp Ser Phe Ala Gly Pro XaaGly Xaa Val Ile Ile Ile Asn 50 55 60 Thr Val Xaa Xaa Val Leu Ser Ala LysVal Ser Cys Gln Arg Lys His 65 70 75 80 His Tyr Tyr Xaa Xaa Lys Gly XaaVal Ser Xaa Leu Arg Thr Ala Phe 85 90 95 Leu Leu Leu Leu Leu Xaa Xaa AlaThr Trp Leu Leu Gly Leu Leu Ala 100 105 110 Val Asn Xaa Asp Xaa Leu SerPhe His Tyr Leu Phe Ala Xaa Phe Ser 115 120 125 Xaa Leu Gln Cys Xaa PheVal Leu Leu Phe His Cys Val Xaa Xaa Xaa 130 135 140 Glu Val Arg Lys HisLeu Xaa Xaa Val Leu Xaa Gly Xaa Lys Leu Xaa 145 150 155 160 Leu Xaa AspSer Ala Thr Thr Arg Ala Thr Leu Leu Thr Arg Ser Leu 165 170 175 Asn CysAsn Xaa Thr Xaa Xaa Xaa Gly Pro Asp Met Leu Arg Thr Xaa 180 185 190 LeuGly Glu Ser Thr Ala Ser Leu Asp Ser Xaa Xaa Arg Asp Glu Gly 195 200 205Xaa Gln Lys Leu Xaa Val Ser Ser Gly Xaa Arg Gly Xaa His Gly Glu 210 215220 Pro Asp Xaa Ser Xaa Xaa Pro Arg Xaa Xaa Lys Xaa Xaa Xaa Gly Xaa 225230 235 240 Asp Ser Asp Ser Asp Ser Glu Leu Ser Leu Asp Glu Xaa Ser SerSer 245 250 255 Tyr Ala Ser Ser His Xaa Ser Asp Ser Glu Asp Asp Gly XaaXaa Ala 260 265 270 Glu Xaa Lys Trp Xaa Pro Ala Xaa Gly Xaa Xaa His SerThr Pro Lys 275 280 285 Xaa Asp Ala Xaa Ala Asn His Val Pro Ala Gly TrpPro Asp Xaa Ser 290 295 300 Leu Ala Xaa Ser Asp Ser Glu Xaa Xaa Xaa XaaXaa Pro Xaa Leu Lys 305 310 315 320 Val Glu Thr Lys Val Ser Val Glu LeuHis Arg Xaa Xaa Gln Gly Xaa 325 330 335 His Xaa Gly Xaa Xaa Pro Xaa AspXaa Glu Ser Gly Xaa Xaa Ala Xaa 340 345 350 Xaa Xaa Xaa Xaa Xaa Ser SerGln Pro Xaa Glu Gln Arg Lys Gly Ile 355 360 365 Leu Lys Asn Lys Val ThrTyr Pro Pro Pro Leu Xaa Xaa Xaa Glu Gln 370 375 380 Xaa Leu Lys Xaa ArgLeu Arg Glu Lys Leu Ala Asp Cys Glu Gln Ser 385 390 395 400 Pro Thr SerSer Arg Thr Ser Ser Leu Gly Ser Gly Xaa Xaa Xaa Xaa 405 410 415 Xaa XaaAsp Cys Xaa Ile Thr Xaa Lys Xaa Pro Xaa Arg Glu Pro Gly 420 425 430 ArgXaa His Leu Asn Gly Val Ala Met Asn Val Arg Thr Gly Ser Ala 435 440 445Gln Ala Xaa Gly Ser Asp Ser Glu Lys Pro 450 455 29 3313 PRT Rattusnorvegicus 29 Met Ala Arg Arg Pro Leu Trp Trp Gly Leu Pro Gly Pro SerThr Pro 1 5 10 15 Leu Leu Leu Leu Leu Leu Phe Ser Leu Phe Pro Ser SerArg Glu Glu 20 25 30 Met Gly Gly Gly Gly Asp Gln Gly Trp Asp Pro Gly ValAla Thr Ala 35 40 45 Thr Gly Pro Arg Ala Gln Ile Gly Ser Gly Ala Val AlaLeu Cys Pro 50 55 60 Glu Ser Pro Gly Val Trp Glu Asp Gly Asp Pro Gly LeuGly Val Arg 65 70 75 80 Glu Pro Val Phe Met Lys Leu Arg Val Gly Arg GlnAsn Ala Arg Asn 85 90 95 Gly Arg Gly Ala Pro Glu Gln Pro Asn Arg Glu ProVal Val Gln Ala 100 105 110 Leu Gly Ser Arg Glu Gln Glu Ala Gly Gln GlySer Gly Tyr Leu Leu 115 120 125 Cys Trp His Pro Glu Ile Ser Ser Cys GlyArg Thr Gly His Leu Arg 130 135 140 Arg Gly Ser Leu Pro Leu Asp Ala LeuSer Pro Gly Asp Ser Asp Leu 145 150 155 160 Arg Asn Ser Ser Pro His ProSer Glu Leu Leu Ala Gln Pro Asp Ser 165 170 175 Pro Arg Pro Val Ala PheGln Arg Asn Gly Arg Arg Ser Ile Arg Lys 180 185 190 Arg Val Glu Thr PheArg Cys Cys Gly Lys Leu Trp Glu Pro Gly His 195 200 205 Lys Gly Gln GlyGlu Arg Ser Ala Thr Ser Thr Val Asp Arg Gly Pro 210 215 220 Leu Arg ArgAsp Cys Leu Pro Gly Ser Leu Gly Ser Gly Leu Gly Glu 225 230 235 240 AspSer Ala Pro Arg Ala Val Arg Thr Ala Pro Ala Pro Gly Ser Ala 245 250 255Pro His Glu Ser Arg Thr Ala Pro Glu Arg Met Arg Ser Arg Gly Leu 260 265270 Phe Arg Arg Gly Phe Leu Phe Glu Arg Pro Gly Pro Arg Pro Pro Gly 275280 285 Phe Pro Thr Gly Ala Glu Ala Lys Arg Ile Leu Ser Thr Asn Gln Ala290 295 300 Arg Ser Arg Arg Ala Ala Asn Arg His Pro Gln Phe Pro Gln TyrAsn 305 310 315 320 Tyr Gln Thr Leu Val Pro Glu Asn Glu Ala Ala Gly ThrAla Val Leu 325 330 335 Arg Val Val Ala Gln Asp Pro Asp Pro Gly Glu AlaGly Arg Leu Val 340 345 350 Tyr Ser Leu Ala Ala Leu Met Asn Ser Arg SerLeu Glu Leu Phe Ser 355 360 365 Ile Asp Pro Gln Ser Gly Leu Ile Arg ThrAla Ala Ala Leu Asp Arg 370 375 380 Glu Ser Met Glu Arg His Tyr Leu ArgVal Thr Ala Gln Asp His Gly 385 390 395 400 Ser Pro Arg Leu Ser Ala ThrThr Met Val Ala Val Thr Val Ala Asp 405 410 415 Arg Asn Asp His Ala ProVal Phe Glu Gln Ala Gln Tyr Arg Glu Thr 420 425 430 Leu Arg Glu Asn ValGlu Glu Gly Tyr Pro Ile Leu Gln Leu Arg Ala 435 440 445 Thr Asp Gly AspAla Pro Pro Asn Ala Asn Leu Arg Tyr Arg Phe Val 450 455 460 Gly Ser ProAla Ala Arg Thr Ala Ala Ala Ala Ala Phe Glu Ile Asp 465 470 475 480 ProArg Ser Gly Leu Ile Ser Thr Ser Gly Arg Val Asp Arg Glu His 485 490 495Met Glu Ser Tyr Glu Leu Val Val Glu Ala Ser Asp Gln Gly Gln Glu 500 505510 Pro Gly Pro Arg Ser Ala Thr Val Arg Val His Ile Thr Val Leu Asp 515520 525 Glu Asn Asp Asn Ala Pro Gln Phe Ser Glu Lys Arg Tyr Val Ala Gln530 535 540 Val Arg Glu Asp Val Arg Pro His Thr Val Val Leu Arg Val ThrAla 545 550 555 560 Thr Asp Lys Asp Lys Asp Ala Asn Gly Leu Val His TyrAsn Ile Ile 565 570 575 Ser Gly Asn Ser Arg Gly His Phe Ala Ile Asp SerLeu Thr Gly Glu 580 585 590 Ile Gln Val Met Ala Pro Leu Asp Phe Glu AlaGlu Arg Glu Tyr Ala 595 600 605 Leu Arg Ile Arg Ala Gln Asp Ala Gly ArgPro Pro Leu Ser Asn Asn 610 615 620 Thr Gly Leu Ala Ser Ile Gln Val ValAsp Ile Asn Asp His Ser Pro 625 630 635 640 Ile Phe Val Ser Thr Pro PheGln Val Ser Val Leu Glu Asn Ala Pro 645 650 655 Leu Gly His Ser Val IleHis Ile Gln Ala Val Asp Ala Asp His Gly 660 665 670 Glu Asn Ser Arg LeuGlu Tyr Ser Leu Thr Gly Val Ala Ser Asp Thr 675 680 685 Pro Phe Val IleAsn Ser Ala Thr Gly Trp Val Ser Val Ser Gly Pro 690 695 700 Leu Asp ArgGlu Ser Val Glu His Tyr Phe Phe Gly Val Glu Ala Arg 705 710 715 720 AspHis Gly Ser Pro Pro Leu Ser Ala Ser Ala Ser Val Thr Val Thr 725 730 735Val Leu Asp Val Asn Asp Asn Arg Pro Glu Phe Thr Met Lys Glu Tyr 740 745750 His Leu Arg Leu Asn Glu Asp Ala Ala Val Gly Thr Ser Val Val Ser 755760 765 Val Thr Ala Val Asp Arg Asp Ala Asn Ser Ala Ile Ser Tyr Gln Ile770 775 780 Thr Gly Gly Asn Thr Arg Asn Arg Phe Ala Ile Ser Thr Gln GlyGly 785 790 795 800 Met Gly Leu Val Thr Leu Ala Leu Pro Leu Asp Tyr LysGln Glu Arg 805 810 815 Tyr Phe Lys Leu Val Leu Thr Ala Ser Asp Arg AlaLeu His Asp His 820 825 830 Cys Tyr Val His Ile Asn Ile Thr Asp Ala AsnThr His Arg Pro Val 835 840 845 Phe Gln Ser Ala His Tyr Ser Val Ser MetAsn Glu Asp Arg Pro Val 850 855 860 Gly Ser Thr Val Val Val Ile Ser AlaSer Asp Asp Asp Val Gly Glu 865 870 875 880 Asn Ala Arg Ile Thr Tyr LeuLeu Glu Asp Asn Leu Pro Gln Phe Arg 885 890 895 Ile Asp Ala Asp Ser GlyAla Ile Thr Leu Gln Ala Pro Leu Asp Tyr 900 905 910 Glu Asp Gln Val ThrTyr Thr Leu Ala Ile Thr Ala Arg Asp Asn Gly 915 920 925 Ile Pro Gln LysAla Asp Thr Thr Tyr Val Glu Val Met Val Asn Asp 930 935 940 Val Asn AspAsn Ala Pro Gln Phe Val Ala Ser His Tyr Thr Gly Leu 945 950 955 960 ValSer Glu Asp Ala Pro Pro Phe Thr Ser Val Leu Gln Ile Ser Ala 965 970 975Thr Asp Arg Asp Ala His Ala Asn Gly Arg Val Gln Tyr Thr Phe Gln 980 985990 Asn Gly Glu Asp Gly Asp Gly Asp Phe Thr Ile Glu Pro Thr Ser Gly 9951000 1005 Ile Val Arg Thr Val Arg Arg Leu Asp Arg Glu Ala Val Pro ValTyr 1010 1015 1020 Glu Leu Thr Ala Tyr Ala Val Asp Arg Gly Val Pro ProLeu Arg Thr 1025 1030 1035 1040 Pro Val Ser Ile Gln Val Thr Val Gln AspVal Asn Asp Asn Ala Pro 1045 1050 1055 Val Phe Pro Ala Glu Glu Phe GluVal Arg Val Lys Glu Asn Ser Ile 1060 1065 1070 Val Gly Ser Val Val AlaGln Ile Thr Ala Val Asp Pro Asp Asp Gly 1075 1080 1085 Pro Asn Ala HisIle Met Tyr Gln Ile Val Glu Gly Asn Ile Pro Glu 1090 1095 1100 Leu PheGln Met Asp Ile Phe Ser Gly Glu Leu Thr Ala Leu Ile Asp 1105 1110 11151120 Leu Asp Tyr Glu Ala Arg Gln Glu Tyr Val Ile Val Val Gln Ala Thr1125 1130 1135 Ser Ala Pro Leu Val Ser Arg Ala Thr Val His Val Arg LeuVal Asp 1140 1145 1150 Gln Asn Asp Asn Ser Pro Val Leu Asn Asn Phe GlnIle Leu Phe Asn 1155 1160 1165 Asn Tyr Val Ser Asn Arg Ser Asp Thr PhePro Ser Gly Ile Ile Gly 1170 1175 1180 Arg Ile Pro Ala Tyr Asp Pro AspVal Ser Asp His Leu Phe Tyr Ser 1185 1190 1195 1200 Phe Glu Arg Gly AsnGlu Leu Gln Leu Leu Val Val Asn Gln Thr Ser 1205 1210 1215 Gly Glu LeuArg Leu Ser Arg Lys Leu Asp Asn Asn Arg Pro Leu Val 1220 1225 1230 AlaSer Met Leu Val Thr Val Thr Asp Gly Leu His Ser Val Thr Ala 1235 12401245 Gln Cys Val Leu Arg Val Val Ile Ile Thr Glu Glu Leu Leu Ala Asn1250 1255 1260 Ser Leu Thr Val Arg Leu Glu Asn Met Trp Gln Glu Arg PheLeu Ser 1265 1270 1275 1280 Pro Leu Leu Gly His Phe Leu Glu Gly Val AlaAla Val Leu Ala Thr 1285 1290 1295 Pro Thr Glu Asp Val Phe Ile Phe AsnIle Gln Asn Asp Thr Asp Val 1300 1305 1310 Gly Gly Thr Val Leu Asn ValSer Phe Ser Ala Leu Ala Pro Arg Gly 1315 1320 1325 Ala Gly Ala Gly AlaAla Gly Pro Trp Phe Ser Ser Glu Glu Leu Gln 1330 1335 1340 Glu Gln LeuTyr Val Arg Arg Ala Ala Leu Ala Ala Arg Ser Leu Leu 1345 1350 1355 1360Asp Val Leu Pro Phe Asp Asp Asn Val Cys Leu Arg Glu Pro Cys Glu 13651370 1375 Asn Tyr Met Lys Cys Val Ser Val Leu Arg Phe Asp Ser Ser AlaPro 1380 1385 1390 Phe Leu Ala Ser Ala Ser Thr Leu Phe Arg Pro Ile GlnPro Ile Ala 1395 1400 1405 Gly Leu Arg Cys Arg Cys Pro Pro Gly Phe ThrGly Asp Phe Cys Glu 1410 1415 1420 Thr Glu Leu Asp Leu Cys Tyr Ser AsnPro Cys Arg Asn Gly Gly Ala 1425 1430 1435 1440 Cys Ala Arg Arg Glu GlyGly Tyr Thr Cys Val Cys Arg Pro Arg Phe 1445 1450 1455 Thr Gly Glu AspCys Glu Leu Asp Thr Glu Ala Gly Arg Cys Val Pro 1460 1465 1470 Gly ValCys Arg Asn Gly Gly Thr Cys Thr Asn Ala Pro Asn Gly Gly 1475 1480 1485Phe Arg Cys Gln Cys Pro Ala Gly Gly Ala Phe Glu Gly Pro Arg Cys 14901495 1500 Glu Val Ala Ala Arg Ser Phe Pro Pro Ser Ser Phe Val Met PheArg 1505 1510 1515 1520 Gly Leu Arg Gln Arg Phe His Leu Thr Leu Ser LeuSer Phe Ala Thr 1525 1530 1535 Val Gln Pro Ser Gly Leu Leu Phe Tyr AsnGly Arg Leu Asn Glu Lys 1540 1545 1550 His Asp Phe Leu Ala Leu Glu LeuVal Ala Gly Gln Val Arg Leu Thr 1555 1560 1565 Tyr Ser Thr Gly Glu SerSer Thr Val Val Ser Pro Thr Val Pro Gly 1570 1575 1580 Gly Leu Ser AspGly Gln Trp His Thr Val His Leu Arg Tyr Tyr Asn 1585 1590 1595 1600 LysPro Arg Thr Asp Ala Leu Gly Gly Ala Gln Gly Pro Ser Lys Asp 1605 16101615 Lys Val Ala Val Leu Ser Val Asp Asp Cys Asn Val Ala Val Ala Leu1620 1625 1630 Arg Phe Gly Ala Glu Ile Gly Asn Tyr Ser Cys Ala Ala AlaGly Val 1635 1640 1645 Gln Thr Ser Ser Lys Lys Ser Leu Asp Leu Thr GlyPro Leu Leu Leu 1650 1655 1660 Gly Gly Val Pro Asn Leu Pro Glu Asn PhePro Val Ser Arg Lys Asp 1665 1670 1675 1680 Phe Ile Gly Cys Met Arg AspLeu His Ile Asp Gly Arg Arg Val Asp 1685 1690 1695 Met Ala Ala Phe ValAla Asn Asn Gly Thr Thr Ala Gly Cys Gln Ala 1700 1705 1710 Lys Ser HisPhe Cys Ala Ser Gly Pro Cys Lys Asn Gly Gly Leu Cys 1715 1720 1725 SerGlu Arg Trp Gly Gly Phe Ser Cys Asp Cys Pro Val Gly Phe Gly 1730 17351740 Gly Lys Asp Cys Arg Leu Thr Met Ala His Pro Tyr His Phe Gln Gly1745 1750 1755 1760 Asn Gly Thr Leu Ser Trp Asp Phe Gly Asn Asp Met ProVal Ser Val 1765 1770 1775 Pro Trp Tyr Leu Gly Leu Ser Phe Arg Thr ArgAla Thr Lys Gly Val 1780 1785 1790 Leu Met Gln Val Gln Leu Gly Pro HisSer Val Leu Leu Cys Lys Leu 1795 1800 1805 Asp Gln Gly Leu Leu Ser ValThr Leu Ser Arg Ala Ser Gly His Ala 1810 1815 1820 Val His Leu Leu LeuAsp Gln Met Thr Val Ser Asp Gly Arg Trp His 1825 1830 1835 1840 Asp LeuArg Leu Glu Leu Gln Glu Glu Pro Gly Gly Arg Arg Gly His 1845 1850 1855His Ile Phe Met Val Ser Leu Asp Phe Thr Leu Phe Gln Asp Thr Met 18601865 1870 Ala Met Gly Ser Glu Leu Glu Gly Leu Lys Val Lys His Leu HisVal 1875 1880 1885 Gly Gly Pro Pro Pro Ser Ser Lys Glu Glu Gly Pro GlnGly Leu Val 1890 1895 1900 Gly Cys Ile Gln Gly Val Trp Thr Gly Phe ThrPro Phe Gly Ser Ser 1905 1910 1915 1920 Ala Leu Pro Pro Pro Ser His ArgIle Asn Val Glu Pro Gly Cys Thr 1925 1930 1935 Val Thr Asn Pro Cys AlaSer Gly Pro Cys Pro Pro His Ala Asn Cys 1940 1945 1950 Lys Asp Leu TrpGln Thr Phe Ser Cys Thr Cys Trp Pro Gly Tyr Tyr 1955 1960 1965 Gly ProGly Cys Val Asp Ala Cys Leu Leu Asn Pro Cys Gln Asn Gln 1970 1975 1980Gly Ser Cys Arg His Leu Gln Gly Gly Pro His Gly Tyr Thr Cys Asp 19851990 1995 2000 Cys Ala Ser Gly Tyr Phe Gly Gln His Cys Glu His Arg MetAsp Gln 2005 2010 2015 Gln Cys Pro Arg Gly Trp Trp Gly Ser Pro Thr CysGly Pro Cys Asn 2020 2025 2030 Cys Asp Val His Lys Gly Phe Asp Pro AsnCys Asn Lys Thr Ser Gly 2035 2040 2045 Gln Cys His Cys Lys Glu Phe HisTyr Arg Pro Arg Gly Ser Asp Ser 2050 2055 2060 Cys Leu Pro Cys Asp CysTyr Pro Val Gly Ser Thr Ser Arg Ser Cys 2065 2070 2075 2080 Ala Pro HisSer Gly Gln Cys Pro Cys Arg Pro Gly Ala Leu Gly Arg 2085 2090 2095 GlnCys Asn Ser Cys Asp Ser Pro Phe Ala Glu Val Thr Ala Ser Gly 2100 21052110 Cys Arg Val Leu Tyr Asp Ala Cys Pro Lys Ser Leu Arg Ser Gly Val2115 2120 2125 Trp Trp Pro Gln Thr Lys Phe Gly Val Leu Ala Thr Val ProCys Pro 2130 2135 2140 Arg Gly Ala Leu Gly Leu Arg Gly Thr Gly Ala AlaVal Arg Leu Cys 2145 2150 2155 2160 Asp Glu Asp His Gly Trp Leu Glu ProAsp Phe Phe Asn Cys Thr Ser 2165 2170 2175 Pro Ala Phe Arg Glu Leu SerLeu Leu Leu Asp Gly Leu Glu Leu Asn 2180 2185 2190 Lys Thr Ala Leu AspThr Val Glu Ala Lys Lys Leu Ala Gln Arg Leu 2195 2200 2205 Arg Glu ValThr Gly Gln Thr Asp His Tyr Phe Ser Gln Asp Val Arg 2210 2215 2220 ValThr Ala Arg Leu Leu Ala Tyr Leu Leu Ala Phe Glu Ser His Gln 2225 22302235 2240 Gln Gly Phe Gly Leu Thr Ala Thr Gln Asp Ala His Phe Asn GluAsn 2245 2250 2255 Leu Leu Trp Ala Gly Ser Ala Leu Leu Ala Pro Glu ThrGly Asp Leu 2260 2265 2270 Trp Ala Ala Leu Gly Gln Arg Ala Pro Gly GlySer Pro Gly Ser Ala 2275 2280 2285 Gly Leu Val Arg His Leu Glu Glu TyrAla Ala Thr Leu Ala Arg Asn 2290 2295 2300 Met Asp Leu Thr Tyr Leu AsnPro Val Gly Leu Val Thr Pro Asn Ile 2305 2310 2315 2320 Met Leu Ser IleAsp Arg Met Glu Gln Pro Ser Ser Ser Gln Gly Ala 2325 2330 2335 His ArgTyr Pro Arg Tyr His Ser Asn Leu Phe Arg Gly Gln Asp Ala 2340 2345 2350Trp Asp Pro His Thr His Val Leu Leu Pro Ser Gln Ser Pro Gln Pro 23552360 2365 Ser Pro Ser Glu Val Leu Pro Thr Ser Ser Asn Ala Glu Asn AlaThr 2370 2375 2380 Ala Ser Gly Val Val Ser Pro Pro Ala Pro Leu Glu ProGlu Ser Glu 2385 2390 2395 2400 Pro Gly Ile Ser Ile Val Ile Leu Leu ValTyr Arg Ala Leu Gly Gly 2405 2410 2415 Leu Leu Pro Ala Gln Phe Gln AlaGlu Arg Arg Gly Ala Arg Leu Pro 2420 2425 2430 Gln Asn Pro Val Met AsnSer Pro Val Val Ser Val Ala Val Phe Arg 2435 2440 2445 Gly Arg Asn PheLeu Arg Gly Ala Leu Val Ser Pro Ile Asn Leu Glu 2450 2455 2460 Phe ArgLeu Leu Gln Thr Ala Asn Arg Ser Lys Ala Ile Cys Val Gln 2465 2470 24752480 Trp Asp Pro Pro Gly Pro Ala Asp Gln His Gly Met Trp Thr Ala Arg2485 2490 2495 Asp Cys Glu Leu Val His Arg Asn Gly Ser His Ala Arg CysArg Cys 2500 2505 2510 Ser Arg Thr Gly Thr Phe Gly Val Leu Met Asp AlaSer Pro Arg Glu 2515 2520 2525 Arg Leu Glu Gly Asp Leu Glu Leu Leu AlaVal Phe Thr His Val Val 2530 2535 2540 Val Ala Ala Ser Val Thr Ala LeuVal Leu Thr Ala Ala Val Leu Leu 2545 2550 2555 2560 Ser Leu Arg Ser LeuLys Ser Asn Val Arg Gly Ile His Ala Asn Val 2565 2570 2575 Ala Ala AlaLeu Gly Val Ala Glu Leu Leu Phe Leu Leu Gly Ile His 2580 2585 2590 ArgThr His Asn Gln Leu Leu Cys Thr Val Val Ala Ile Leu Leu His 2595 26002605 Tyr Phe Phe Leu Ser Thr Phe Ala Trp Leu Leu Val Gln Gly Leu His2610 2615 2620 Leu Tyr Arg Met Gln Val Glu Pro Arg Asn Val Asp Arg GlyAla Met 2625 2630 2635 2640 Arg Phe Tyr His Ala Leu Gly Trp Gly Val ProAla Val Leu Leu Gly 2645 2650 2655 Leu Ala Val Gly Leu Asp Pro Glu GlyTyr Gly Asn Pro Asp Phe Cys 2660 2665 2670 Trp Ile Ser Ile His Glu ProLeu Ile Trp Ser Phe Ala Gly Pro Ile 2675 2680 2685 Val Leu Val Ile ValMet Asn Gly Ile Met Phe Leu Leu Ala Ala Arg 2690 2695 2700 Thr Ser CysSer Thr Gly Gln Arg Glu Ala Lys Lys Thr Ser Val Leu 2705 2710 2715 2720Arg Thr Leu Arg Ser Ser Phe Leu Leu Leu Leu Leu Val Ser Ala Ser 27252730 2735 Trp Leu Phe Gly Leu Leu Ala Val Asn His Ser Val Leu Ala PheHis 2740 2745 2750 Tyr Leu His Ala Gly Leu Cys Gly Leu Gln Gly Leu AlaVal Leu Leu 2755 2760 2765 Leu Phe Cys Val Leu Asn Ala Asp Ala Arg AlaAla Trp Thr Pro Ala 2770 2775 2780 Cys Leu Gly Lys Lys Ala Ala Pro GluGlu Thr Arg Pro Ala Pro Gly 2785 2790 2795 2800 Pro Gly Ser Gly Ala TyrAsn Asn Thr Ala Leu Phe Glu Glu Ser Gly 2805 2810 2815 Leu Ile Arg IleThr Leu Gly Ala Ser Thr Val Ser Ser Val Ser Ser 2820 2825 2830 Ala ArgSer Gly Arg Ala Gln Asp Gln Asp Ser Gln Arg Gly Arg Ser 2835 2840 2845Tyr Leu Arg Asp Asn Val Leu Val Arg His Gly Ser Thr Ala Glu His 28502855 2860 Ala Glu His Ser Leu Gln Ala His Ala Gly Pro Thr Asp Leu AspVal 2865 2870 2875 2880 Ala Met Phe His Arg Asp Ala Gly Ala Asp Ser AspSer Asp Ser Asp 2885 2890 2895 Leu Ser Leu Glu Glu Glu Arg Ser Leu SerIle Pro Ser Ser Glu Ser 2900 2905 2910 Glu Asp Asn Gly Arg Thr Arg GlyArg Phe Gln Arg Pro Leu Arg Arg 2915 2920 2925 Ala Ala Gln Ser Glu ArgLeu Leu Ala His Pro Lys Asp Val Asp Gly 2930 2935 2940 Asn Asp Leu LeuSer Tyr Trp Pro Ala Leu Gly Glu Cys Glu Ala Ala 2945 2950 2955 2960 ProCys Ala Leu Gln Ala Trp Gly Ser Glu Arg Arg Leu Gly Leu Asp 2965 29702975 Ser Asn Lys Asp Ala Ala Asn Asn Asn Gln Pro Glu Leu Ala Leu Thr2980 2985 2990 Ser Gly Asp Glu Thr Ser Leu Gly Arg Ala Gln Arg Gln ArgLys Gly 2995 3000 3005 Ile Leu Lys Asn Arg Leu Gln Tyr Pro Leu Val ProGln Thr Arg Gly 3010 3015 3020 Thr Pro Glu Leu Ser Trp Cys Arg Ala AlaThr Leu Gly His Arg Ala 3025 3030 3035 3040 Val Pro Ala Ala Ser Tyr GlyArg Ile Tyr Ala Gly Gly Gly Thr Gly 3045 3050 3055 Ser Leu Ser Gln ProAla Ser Arg Tyr Ser Ser Arg Glu Gln Leu Asp 3060 3065 3070 Leu Leu LeuArg Arg Gln Leu Ser Arg Glu Arg Leu Glu Glu Val Pro 3075 3080 3085 ValPro Ala Pro Val Leu His Pro Leu Ser Arg Pro Gly Ser Gln Glu 3090 30953100 Arg Leu Asp Thr Ala Pro Ala Arg Leu Glu Pro Arg Asp Arg Gly Ser3105 3110 3115 3120 Thr Leu Pro Arg Arg Gln Pro Pro Arg Asp Tyr Pro GlyThr Met Ala 3125 3130 3135 Gly Arg Phe Gly Ser Arg Asp Ala Leu Asp LeuGly Ala Pro Arg Glu 3140 3145 3150 Trp Leu Ser Thr Leu Pro Pro Pro ArgArg Asn Arg Asp Leu Asp Pro 3155 3160 3165 Gln His Pro Pro Leu Pro LeuSer Pro Gln Arg Pro Leu Ser Arg Asp 3170 3175 3180 Pro Leu Leu Pro SerArg Pro Leu Asp Ser Leu Ser Arg Ile Ser Asn 3185 3190 3195 3200 Ser ArgGlu Arg Leu Asp Gln Val Pro Ser Arg His Pro Ser Arg Glu 3205 3210 3215Ala Leu Gly Pro Ala Pro Gln Leu Leu Arg Ala Arg Glu Asp Pro Ala 32203225 3230 Ser Gly Pro Ser His Gly Pro Ser Thr Glu Gln Leu Asp Ile LeuSer 3235 3240 3245 Ser Ile Leu Ala Ser Phe Asn Ser Ser Ala Leu Ser SerVal Gln Ser 3250 3255 3260 Ser Ser Thr Pro Ser Gly Pro His Thr Thr AlaThr Pro Ser Ala Thr 3265 3270 3275 3280 Ala Ser Ala Leu Gly Pro Ser ThrPro Arg Ser Ala Thr Ser His Ser 3285 3290 3295 Ile Ser Glu Leu Ser ProAsp Ser Glu Val Pro Arg Ser Glu Gly His 3300 3305 3310 Ser 30 3034 PRTMus musculus 30 Met Ala Pro Ser Ser Pro Arg Val Leu Pro Ala Leu Val LeuLeu Ala 1 5 10 15 Ala Ala Ala Leu Pro Ala Leu Glu Leu Gly Ala Ala AlaTrp Glu Leu 20 25 30 Arg Val Pro Gly Gly Ala Arg Ala Phe Ala Leu Gly ProGly Trp Ser 35 40 45 Tyr Arg Leu Asp Thr Thr Arg Thr Pro Arg Glu Leu LeuAsp Val Ser 50 55 60 Arg Glu Gly Pro Ala Ala Gly Arg Arg Leu Gly Leu GlyAla Gly Thr 65 70 75 80 Leu Gly Cys Ala Arg Leu Ala Gly Arg Leu Leu ProLeu Gln Val Arg 85 90 95 Leu Val Ala Arg Gly Ala Pro Thr Ala Pro Ser LeuVal Leu Arg Ala 100 105 110 Arg Ala Tyr Gly Ala Arg Cys Gly Val Arg LeuLeu Arg Arg Ser Ala 115 120 125 Arg Gly Ala Glu Leu Arg Ser Pro Ala ValArg Ser Val Pro Gly Leu 130 135 140 Gly Asp Ala Leu Cys Phe Pro Ala AlaGly Gly Gly Ala Ala Ser Leu 145 150 155 160 Thr Ser Val Leu Glu Ala IleThr Asn Phe Pro Ala Cys Ser Cys Pro 165 170 175 Pro Val Ala Gly Thr GlyCys Arg Arg Gly Pro Ile Cys Leu Arg Pro 180 185 190 Gly Gly Ser Ala GluLeu Arg Leu Val Cys Ala Leu Gly Arg Ala Ala 195 200 205 Gly Ala Val TrpVal Glu Leu Val Ile Gln Ala Thr Ser Gly Thr Pro 210 215 220 Ser Glu SerPro Ser Val Ser Pro Ser Leu Leu Asn Leu Ser Gln Pro 225 230 235 240 ArgAla Gly Val Val Arg Arg Ser Arg Arg Gly Thr Gly Ser Ser Thr 245 250 255Ser Pro Gln Phe Pro Leu Pro Ser Tyr Gln Val Ser Val Pro Glu Asn 260 265270 Glu Pro Ala Gly Thr Ala Val Ile Glu Leu Arg Ala His Asp Pro Asp 275280 285 Glu Gly Asp Ala Gly Arg Leu Ser Tyr Gln Met Glu Ala Leu Phe Asp290 295 300 Glu Arg Ser Asn Gly Tyr Phe Leu Ile Asp Ala Ala Thr Gly AlaVal 305 310 315 320 Thr Thr Ala Arg Ser Leu Asp Arg Glu Thr Lys Asp ThrHis Val Leu 325 330 335 Lys Val Ser Ala Val Asp His Gly Ser Pro Arg ArgSer Ala Ala Thr 340 345 350 Tyr Leu Thr Val Thr Val Ser Asp Thr Asn AspHis Ser Pro Val Phe 355 360 365 Glu Gln Ser Glu Tyr Arg Glu Arg Ile ArgGlu Asn Leu Glu Val Gly 370 375 380 Tyr Glu Val Leu Thr Ile Arg Ala ThrAsp Gly Asp Ala Pro Ser Asn 385 390 395 400 Ala Asn Met Arg Tyr Arg LeuLeu Glu Gly Ala Gly Gly Val Phe Glu 405 410 415 Ile Asp Ala Arg Ser GlyVal Val Arg Thr Arg Ala Val Val Asp Arg 420 425 430 Glu Glu Ala Ala GluTyr Gln Leu Leu Val Glu Ala Asn Asp Gln Gly 435 440 445 Arg Asn Pro GlyPro Leu Ser Ala Ser Ala Thr Val His Ile Val Val 450 455 460 Glu Asp GluAsn Asp Asn Tyr Pro Gln Phe Ser Glu Lys Arg Tyr Val 465 470 475 480 ValGln Val Pro Glu Asp Val Ala Val Asn Thr Ala Val Leu Arg Val 485 490 495Gln Ala Thr Asp Arg Asp Gln Gly Gln Asn Ala Ala Ile His Tyr Ser 500 505510 Ile Val Ser Gly Asn Leu Lys Gly Gln Phe Tyr Leu His Ser Leu Ser 515520 525 Gly Ser Leu Asp Val Ile Asn Pro Leu Asp Phe Glu Ala Ile Arg Glu530 535 540 Tyr Thr Leu Arg Ile Lys Ala Gln Asp Gly Gly Arg Pro Pro LeuIle 545 550 555 560 Asn Ser Ser Gly Leu Val Ser Val Gln Val Leu Asp ValAsn Asp Asn 565 570 575 Ala Pro Ile Phe Val Ser Ser Pro Phe Gln Ala AlaVal Leu Glu Asn 580 585 590 Val Pro Leu Gly His Ser Val Leu His Ile GlnAla Val Asp Ala Asp 595 600 605 Ala Gly Glu Asn Ala Arg Leu Gln Tyr ArgLeu Val Asp Thr Ala Ser 610 615 620 Thr Ile Val Gly Gly Ser Ser Val AspSer Glu Asn Pro Ala Ser Ala 625 630 635 640 Pro Asp Phe Pro Phe Gln IleHis Asn Ser Ser Gly Trp Ile Thr Val 645 650 655 Cys Ala Glu Leu Asp ArgGlu Glu Val Glu His Tyr Ser Phe Gly Val 660 665 670 Glu Ala Val Asp HisGly Ser Pro Ala Met Ser Ser Ser Ala Ser Val 675 680 685 Ser Ile Thr ValLeu Asp Val Asn Asp Asn Asp Pro Met Phe Thr Gln 690 695 700 Pro Val TyrGlu Leu Arg Leu Asn Glu Asp Ala Ala Val Gly Ser Ser 705 710 715 720 ValLeu Thr Leu Arg Ala Arg Asp Arg Asp Ala Asn Ser Val Ile Thr 725 730 735Tyr Gln Leu Thr Gly Gly Asn Thr Arg Asn Arg Phe Ala Leu Ser Ser 740 745750 Gln Ser Gly Gly Gly Leu Ile Thr Leu Ala Leu Pro Leu Asp Tyr Lys 755760 765 Gln Glu Arg Gln Tyr Val Leu Ala Val Thr Ala Ser Asp Gly Thr Arg770 775 780 Ser His Thr Ala Gln Val Phe Ile Asn Val Thr Asp Ala Asn ThrHis 785 790 795 800 Arg Pro Val Phe Gln Ser Ser His Tyr Thr Val Ser ValSer Glu Asp 805 810 815 Arg Pro Val Gly Thr Ser Ile Ala Thr Ile Ser AlaThr Asp Glu Asp 820 825 830 Thr Gly Glu Asn Ala Arg Ile Thr Tyr Val LeuGlu Asp Pro Val Pro 835 840 845 Gln Phe Arg Ile Asp Pro Asp Thr Gly ThrIle Tyr Thr Met Thr Glu 850 855 860 Leu Asp Tyr Glu Asp Gln Ala Ala TyrThr Leu Ala Ile Thr Ala Gln 865 870 875 880 Asp Asn Gly Ile Pro Gln LysSer Asp Thr Thr Ser Leu Glu Ile Leu 885 890 895 Ile Leu Asp Ala Asn AspAsn Ala Pro Arg Phe Leu Arg Asp Phe Tyr 900 905 910 Gln Gly Ser Val PheGlu Asp Ala Pro Pro Ser Thr Ser Val Leu Gln 915 920 925 Val Ser Ala ThrAsp Arg Asp Ser Gly Pro Asn Gly Arg Leu Leu Tyr 930 935 940 Thr Phe GlnGly Gly Asp Asp Gly Asp Gly Asp Phe Tyr Ile Glu Pro 945 950 955 960 ThrSer Gly Val Ile Arg Thr Gln Arg Arg Leu Asp Arg Glu Asn Val 965 970 975Ala Val Tyr Asn Leu Trp Ala Leu Ala Val Asp Arg Gly Ser Pro Asn 980 985990 Pro Leu Ser Ala Ser Val Gly Ile Gln Val Ser Val Leu Asp Ile Asn 9951000 1005 Asp Asn Pro Pro Val Phe Glu Lys Asp Glu Leu Glu Leu Phe ValGlu 1010 1015 1020 Glu Asn Ser Pro Val Gly Ser Val Val Ala Arg Ile ArgAla Asn Asp 1025 1030 1035 1040 Pro Asp Glu Gly Pro Asn Ala Gln Ile IleTyr Gln Ile Val Glu Gly 1045 1050 1055 Asn Val Pro Glu Val Phe Gln LeuAsp Leu Leu Ser Gly Asp Leu Arg 1060 1065 1070 Ala Leu Val Glu Leu AspPhe Glu Val Arg Arg Asp Tyr Met Leu Val 1075 1080 1085 Val Gln Ala ThrSer Ala Pro Leu Val Ser Arg Ala Thr Val His Ile 1090 1095 1100 Arg LeuLeu Asp Gln Asn Asp Asn Pro Pro Glu Leu Pro Asp Phe Gln 1105 1110 11151120 Ile Leu Phe Asn Asn Tyr Val Thr Asn Lys Ser Asn Ser Phe Pro Ser1125 1130 1135 Gly Val Ile Gly Arg Ile Pro Ala His Asp Pro Asp Leu SerAsp Ser 1140 1145 1150 Leu Asn Tyr Thr Phe Leu Gln Gly Asn Glu Leu SerLeu Leu Leu Leu 1155 1160 1165 Asp Pro Ala Thr Gly Glu Leu Gln Leu SerArg Asp Leu Asp Asn Asn 1170 1175 1180 Arg Pro Leu Glu Ala Leu Met GluVal Ser Val Ser Asp Gly Ile His 1185 1190 1195 1200 Ser Val Thr Ala LeuCys Thr Leu Arg Val Thr Ile Ile Thr Asp Asp 1205 1210 1215 Met Leu ThrAsn Ser Ile Thr Val Arg Leu Glu Asn Met Ser Gln Glu 1220 1225 1230 LysPhe Leu Ser Pro Leu Leu Ser Leu Phe Val Glu Gly Val Ala Thr 1235 12401245 Val Leu Ser Thr Thr Lys Asp Asp Ile Phe Val Phe Asn Ile Gln Asn1250 1255 1260 Asp Thr Asp Val Ser Ser Asn Ile Leu Asn Val Thr Phe SerAla Leu 1265 1270 1275 1280 Leu Pro Gly Gly Thr Arg Gly Arg Phe Phe ProSer Glu Asp Leu Gln 1285 1290 1295 Glu Gln Ile Tyr Leu Asn Arg Thr LeuLeu Thr Thr Ile Ser Ala Gln 1300 1305 1310 Arg Val Leu Pro Phe Asp AspAsn Ile Cys Leu Arg Glu Pro Cys Glu 1315 1320 1325 Asn Tyr Met Lys CysVal Ser Val Leu Arg Phe Asp Ser Ser Ala Pro 1330 1335 1340 Phe Ile SerSer Thr Thr Val Leu Phe Arg Pro Ile His Pro Ile Thr 1345 1350 1355 1360Gly Leu Arg Cys Arg Cys Pro Pro Gly Phe Thr Gly Asp Tyr Cys Glu 13651370 1375 Thr Glu Ile Asp Leu Cys Tyr Ser Asn Pro Cys Gly Ala Asn GlyArg 1380 1385 1390 Cys Arg Ser Arg Glu Gly Gly Tyr Thr Cys Glu Cys PheGlu Asp Phe 1395 1400 1405 Thr Gly Glu His Cys Gln Val Asn Val Arg SerGly Arg Cys Ala Ser 1410 1415 1420 Gly Val Cys Lys Asn Gly Gly Thr CysVal Asn Leu Leu Ile Gly Gly 1425 1430 1435 1440 Phe His Cys Val Cys ProPro Gly Glu Tyr Glu His Pro Tyr Cys Glu 1445 1450 1455 Val Ser Thr ArgSer Phe Pro Pro Gln Ser Phe Val Thr Phe Arg Gly 1460 1465 1470 Leu ArgGln Arg Phe His Phe Thr Val Ser Leu Ala Phe Ala Thr Gln 1475 1480 1485Asp Arg Asn Ala Leu Leu Leu Tyr Asn Gly Arg Phe Asn Glu Lys His 14901495 1500 Asp Phe Ile Ala Leu Glu Ile Val Glu Glu Gln Leu Gln Leu ThrPhe 1505 1510 1515 1520 Ser Ala Gly Glu Thr Thr Thr Thr Val Thr Pro GlnVal Pro Gly Gly 1525 1530 1535 Val Ser Asp Gly Arg Trp His Ser Val LeuVal Gln Tyr Tyr Asn Lys 1540 1545 1550 Pro Asn Ile Gly His Leu Gly LeuPro His Gly Pro Ser Gly Glu Lys 1555 1560 1565 Val Ala Val Val Thr ValAsp Asp Cys Asp Ala Ala Val Ala Val His 1570 1575 1580 Phe Gly Ser TyrVal Gly Asn Tyr Ser Cys Ala Ala Gln Gly Thr Gln 1585 1590 1595 1600 SerGly Ser Lys Lys Ser Leu Asp Leu Thr Gly Pro Leu Leu Leu Gly 1605 16101615 Gly Val Pro Asn Leu Pro Glu Asp Phe Pro Val His Ser Arg Gln Phe1620 1625 1630 Val Gly Cys Met Arg Asn Leu Ser Ile Asp Gly Arg Ile ValAsp Met 1635 1640 1645 Ala Ala Phe Ile Ala Asn Asn Gly Thr Arg Ala GlyCys Ala Ser Gln 1650 1655 1660 Arg Asn Phe Cys Asp Gly Thr Ser Cys GlnAsn Gly Gly Thr Cys Val 1665 1670 1675 1680 Asn Arg Trp Asn Thr Tyr LeuCys Glu Cys Pro Leu Arg Phe Gly Gly 1685 1690 1695 Lys Asn Cys Glu GlnAla Met Pro His Pro Gln Arg Phe Thr Gly Glu 1700 1705 1710 Ser Val ValLeu Trp Ser Asp Leu Asp Ile Thr Ile Ser Val Pro Trp 1715 1720 1725 TyrLeu Gly Leu Met Phe Arg Thr Arg Lys Glu Asp Gly Val Leu Met 1730 17351740 Glu Ala Thr Ala Gly Thr Ser Ser Arg Leu His Leu Gln Ile Leu Asn1745 1750 1755 1760 Ser Tyr Ile Arg Phe Glu Val Ser Tyr Gly Pro Ser AspVal Ala Ser 1765 1770 1775 Met Gln Leu Ser Lys Ser Arg Ile Thr Asp GlyGly Trp His His Leu 1780 1785 1790 Leu Ile Glu Leu Arg Ser Ala Lys GluGly Lys Asp Ile Lys Tyr Leu 1795 1800 1805 Ala Val Met Thr Leu Asp TyrGly Met Asp Gln Ser Thr Val Gln Ile 1810 1815 1820 Gly Asn Gln Leu ProGly Leu Lys Met Arg Thr Ile Val Ile Gly Gly 1825 1830 1835 1840 Val ThrGlu Asp Lys Val Ser Val Arg His Gly Phe Arg Gly Cys Met 1845 1850 1855Gln Gly Val Arg Met Gly Glu Thr Ser Thr Asn Ile Ala Thr Leu Asn 18601865 1870 Met Asn Asp Ala Leu Lys Val Arg Val Lys Asp Gly Cys Asp ValGlu 1875 1880 1885 Asp Pro Cys Ala Ser Ser Pro Cys Pro Pro His Arg ProCys Arg Asp 1890 1895 1900 Thr Trp Asp Ser Tyr Ser Cys Ile Cys Asp ArgGly Tyr Phe Gly Lys 1905 1910 1915 1920 Lys Cys Val Asp Ala Cys Leu LeuAsn Pro Cys Lys His Val Ala Ala 1925 1930 1935 Cys Val Arg Ser Pro AsnThr Pro Arg Gly Tyr Ser Cys Glu Cys Gly 1940 1945 1950 Pro Gly His TyrGly Gln Tyr Cys Glu Asn Lys Val Asp Leu Pro Cys 1955 1960 1965 Pro LysGly Trp Trp Gly Asn Pro Val Cys Gly Pro Cys His Cys Ala 1970 1975 1980Val Ser Gln Gly Phe Asp Pro Asp Cys Asn Lys Thr Asn Gly Gln Cys 19851990 1995 2000 Gln Cys Lys Glu Asn Tyr Tyr Lys Pro Pro Ala Gln Asp AlaCys Leu 2005 2010 2015 Pro Cys Asp Cys Phe Pro His Gly Ser His Ser ArgAla Cys Asp Met 2020 2025 2030 Asp Thr Gly Gln Cys Ala Cys Lys Pro GlyVal Ile Gly Arg Gln Cys 2035 2040 2045 Asn Arg Cys Asp Asn Pro Phe AlaGlu Val Thr Ser Leu Gly Cys Glu 2050 2055 2060 Val Ile Tyr Asn Gly CysPro Arg Ala Phe Glu Ala Gly Ile Trp Trp 2065 2070 2075 2080 Pro Gln ThrLys Phe Gly Gln Pro Ala Ala Val Pro Cys Pro Lys Gly 2085 2090 2095 SerVal Gly Asn Ala Val Arg His Cys Ser Gly Glu Lys Gly Trp Leu 2100 21052110 Pro Pro Glu Leu Phe Asn Cys Thr Ser Gly Ser Phe Val Asp Leu Lys2115 2120 2125 Ala Leu Asn Glu Lys Leu Asn Arg Asn Glu Thr Arg Met AspGly Asn 2130 2135 2140 Arg Ser Leu Arg Leu Ala Lys Ala Leu Arg Asn AlaThr Gln Gly Asn 2145 2150 2155 2160 Ser Thr Leu Phe Gly Asn Asp Val ArgThr Ala Tyr Gln Leu Leu Ala 2165 2170 2175 Arg Ile Leu Gln His Glu SerArg Gln Gln Gly Phe Asp Leu Ala Ala 2180 2185 2190 Thr Arg Glu Ala AsnPhe His Glu Asp Val Val His Thr Gly Ser Ala 2195 2200 2205 Leu Leu AlaPro Ala Thr Glu Ala Ser Trp Glu Gln Ile Gln Arg Ser 2210 2215 2220 GluAla Gly Ala Ala Gln Leu Leu Arg His Phe Glu Ala Tyr Phe Ser 2225 22302235 2240 Asn Val Ala Arg Asn Val Lys Arg Thr Tyr Leu Arg Pro Phe ValIle 2245 2250 2255 Val Thr Ala Asn Met Ile Leu Ala Val Asp Ile Phe AspLys Leu Asn 2260 2265 2270 Phe Thr Gly Ala Gln Val Pro Arg Phe Glu AspIle Gln Glu Glu Leu 2275 2280 2285 Pro Arg Glu Leu Glu Ser Ser Val SerPhe Pro Ala Asp Thr Phe Lys 2290 2295 2300 Pro Pro Glu Lys Lys Glu GlyPro Val Val Arg Leu Thr Asn Arg Arg 2305 2310 2315 2320 Thr Thr Pro LeuThr Ala Gln Pro Glu Pro Arg Ala Glu Arg Glu Thr 2325 2330 2335 Ser SerSer Arg Arg Arg Arg His Pro Asp Glu Pro Gly Gln Phe Ala 2340 2345 2350Val Ala Leu Val Val Ile Tyr Arg Thr Leu Gly Gln Leu Leu Pro Glu 23552360 2365 His Tyr Asp Pro Asp His Arg Ser Leu Arg Leu Pro Asn Arg ProVal 2370 2375 2380 Ile Asn Thr Pro Val Val Ser Ala Met Val Tyr Ser GluGly Thr Pro 2385 2390 2395 2400 Leu Pro Ser Ser Leu Gln Arg Pro Ile LeuVal Glu Phe Ser Leu Leu 2405 2410 2415 Glu Thr Glu Glu Arg Ser Lys ProVal Cys Val Phe Trp Asn His Ser 2420 2425 2430 Leu Asp Thr Gly Gly ThrGly Gly Trp Ser Ala Lys Gly Cys Glu Leu 2435 2440 2445 Leu Ser Arg AsnArg Thr His Val Thr Cys Gln Cys Ser His Ser Ala 2450 2455 2460 Ser CysAla Val Leu Met Asp Ile Ser Arg Arg Glu His Gly Glu Val 2465 2470 24752480 Leu Pro Leu Lys Ile Ile Thr Tyr Ala Ala Leu Ser Leu Ser Leu Val2485 2490 2495 Ala Leu Leu Val Ala Phe Val Leu Leu Ser Leu Val Arg ThrLeu Arg 2500 2505 2510 Ser Asn Leu His Ser Ile His Lys Asn Leu Ile AlaAla Leu Phe Phe 2515 2520 2525 Ser Gln Leu Ile Phe Met Val Gly Ile AsnGln Thr Glu Asn Pro Phe 2530 2535 2540 Leu Cys Thr Val Val Ala Ile LeuLeu His Tyr Val Ser Met Gly Thr 2545 2550 2555 2560 Phe Ala Trp Thr LeuVal Glu Asn Leu His Val Tyr Arg Met Leu Thr 2565 2570 2575 Glu Val ArgAsn Ile Asp Thr Gly Pro Met Arg Phe Tyr His Val Val 2580 2585 2590 GlyTrp Gly Ile Pro Ala Ile Val Thr Gly Leu Ala Val Gly Leu Asp 2595 26002605 Pro Gln Gly Tyr Gly Asn Pro Asp Phe Cys Trp Leu Ser Leu Gln Asp2610 2615 2620 Thr Leu Ile Trp Ser Phe Ala Gly Pro Val Gly Thr Val IleIle Ile 2625 2630 2635 2640 Asn Thr Val Ile Phe Val Leu Ser Ala Lys ValSer Cys Gln Arg Lys 2645 2650 2655 His His Tyr Tyr Glu Arg Lys Gly ValVal Ser Met Leu Arg Thr Ala 2660 2665 2670 Phe Leu Leu Leu Leu Leu ValThr Ala Thr Trp Leu Leu Gly Leu Leu 2675 2680 2685 Ala Val Asn Ser AspThr Leu Ser Phe His Tyr Leu Phe Ala Ala Phe 2690 2695 2700 Ser Cys LeuGln Gly Ile Phe Val Leu Leu Phe His Cys Val Ala His 2705 2710 2715 2720Arg Glu Val Arg Lys His Leu Arg Ala Val Leu Ala Gly Lys Lys Leu 27252730 2735 Gln Leu Asp Asp Ser Ala Thr Thr Arg Ala Thr Leu Leu Thr ArgSer 2740 2745 2750 Leu Asn Cys Asn Asn Thr Tyr Ser Glu Gly Pro Asp MetLeu Arg Thr 2755 2760 2765 Ala Leu Gly Glu Ser Thr Ala Ser Leu Asp SerThr Thr Arg Asp Glu 2770 2775 2780 Gly Val Gln Lys Leu Ser Val Ser SerGly Pro Ala Arg Gly Asn His 2785 2790 2795 2800 Gly Glu Pro Asp Thr SerPhe Ile Pro Arg Asn Ser Lys Lys Ala His 2805 2810 2815 Gly Pro Asp SerAsp Ser Asp Ser Glu Leu Ser Leu Asp Glu His Ser 2820 2825 2830 Ser SerTyr Ala Ser Ser His Thr Ser Asp Ser Glu Asp Asp Gly Gly 2835 2840 2845Glu Ala Glu Asp Lys Trp Asn Pro Ala Gly Gly Pro Ala His Ser Thr 28502855 2860 Pro Lys Ala Asp Ala Leu Ala Asn His Val Pro Ala Gly Trp ProAsp 2865 2870 2875 2880 Glu Ser Leu Ala Gly Ser Asp Ser Glu Glu Leu AspThr Glu Pro His 2885 2890 2895 Leu Lys Val Glu Thr Lys Val Ser Val GluLeu His Arg Gln Ala Gln 2900 2905 2910 Gly Asn His Cys Gly Asp Arg ProSer Asp Pro Glu Ser Gly Val Leu 2915 2920 2925 Ala Lys Pro Val Ala ValLeu Ser Ser Gln Pro Gln Glu Gln Arg Lys 2930 2935 2940 Gly Ile Leu LysAsn Lys Val Thr Tyr Pro Pro Pro Leu Pro Glu Gln 2945 2950 2955 2960 ProLeu Lys Ser Arg Leu Arg Glu Lys Leu Ala Asp Cys Glu Gln Ser 2965 29702975 Pro Thr Ser Ser Arg Thr Ser Ser Leu Gly Ser Gly Asp Gly Val His2980 2985 2990 Ala Thr Asp Cys Val Ile Thr Ile Lys Thr Pro Arg Arg GluPro Gly 2995 3000 3005 Arg Glu His Leu Asn Gly Val Ala Met Asn Val ArgThr Gly Ser Ala 3010 3015 3020 Gln Ala Asn Gly Ser Asp Ser Glu Lys Pro3025 3030 31 1072 DNA Homo sapiens 31 ccctgcccct aggttcctgg ccaacacgtccttccagggc cgcacgggcc ccgtgtgggt 60 gacaggcagc tcccaggtac acatgtctcggcactttaag gtgtggagcc ttcgccggga 120 cccacggggc gccccggcct gggccacggtgggcagctgg cgggacggcc agctggactt 180 ggaaccggga ggtgcctctg cacggcccccgcccccacag ggtgcccagg tctggcccaa 240 gctgcgtgtg gtaacgctgt tggaacacccatttgtgttt gcccgtgatc cagacgaaga 300 cgggcagtgc ccagcggggc agctgtgcctggaccctggc accaacgact cggccaccct 360 ggacgcactg ttcgccgcgc tggccaacggctcagcgccc cgtgccctgc gcaagtgctg 420 ctacggctac tgcattgacc tgctggagcggctggcggag gacacgccct tcgacttcga 480 gctgtacctc gtgggtgacg gcaagtacggcgccctgcgg gacggccgct ggaccggcct 540 ggtcggggac ctgctggccg gccgggcccacatggcggtc accagcttca gtatcaactc 600 cgcccgctca caggtggtgg acttcaccagccccttcttc tccaccagcc tgggcatcat 660 ggtgcgggca cgggacacgg cctcacccatcggtgccttt atgtggcccc tgcactggtc 720 cacgtggctg ggcgtctttg cggccctgcacctcaccgcg ctcttcctca ccgtgtacga 780 gtggcgtagc ccctacggcc tcacgccacgtggccgcaac cgcagcaccg tcttctccta 840 ctcctcagcc ctcaacctgt gctacgccatcctcttcaga cgcaccgtgt ccagcaagac 900 gcccaagtgc cccacgggcc gcctgctcatgaacctctgg gccatcttct gcctgctggt 960 gctgtccagc tacacggcca acctggctgccgtcatggtc ggggacaaga ccttcgagga 1020 gctgtcgggg atccacgacc ccaaggtgggcggcctcggg gggctgcggg tg 1072 32 1083 DNA Artificial Sequence ConsensusSequence 32 cnctgccncc nangnncctg nccancncgn nnctnccann nnngcnncnggcncngngng 60 nggtgncang ancnccncng gnanncnngn cncnncncnt nannntgtgnancnnntngc 120 cnggncncnn gggnngnncn gnccnggnnc ancngnngnn agcnnncnnnncngccnngn 180 tnnactnnng gnaccngnag nngncnnngc ncgngncccc gnnncnncnnnntgncnngn 240 gtnnctggcc naancnncgt nntnnnanng cngnnngnnc ncccntntgngtnnnngncn 300 gnnnccagac gaagacgggc agtgcccagc ggggcagctg tgcctggaccctggcaccaa 360 cgactcggcc accctggacg cactgttcgc cgcgctggcc aacggctcagcgccccgtgc 420 cctgcgcaag tgctgctacg gctactgcat tgacctgctg gagcggctggcggaggacac 480 gcccttcgac ttcgagctgt acctcgtggg tgacggcaag tacggcgccctgcgggacgg 540 ccgctggacc ggcctggtcg gggacctgct ggccggccgg gcccacatggcggtcaccag 600 cttcagtatc aactccgccc gctcacaggt ggtggacttc accagccccttcttctccac 660 cagcctgggc atcatggtgc gggcacggga cacggcctca cccatcggtgcctttatgtg 720 gcccctgcac tggtccacgt ggctgggcgt ctttgcggcc ctgcacctcaccgcgctctt 780 cctcaccgtg tacgagtggc gtagccccta cggcctcacg ccacgtggccgcaaccgcag 840 caccgtcttc tcctactcct cagccctcaa cctgtgctac gccatcctcttcagacgcac 900 cgtgtccagc aagacgccca agtgccccac gggccgcctg ctcatgaacctctgggccat 960 cttctgcctg ctggtgctgt ccagctacac ggccaacctg gctgccgtcatggtcgggga 1020 caagaccttc gaggagctgt cggggatcca cgaccccaag gnnnncngcntcggntgngg 1080 gng 1083 33 901 PRT Homo sapiens 33 Met Glu Phe Val ArgAla Leu Trp Leu Gly Leu Ala Leu Ala Leu Gly 1 5 10 15 Pro Gly Ser AlaGly Gly His Pro Gln Pro Cys Gly Val Leu Ala Arg 20 25 30 Leu Gly Gly SerVal Arg Leu Gly Ala Leu Leu Pro Arg Ala Pro Leu 35 40 45 Ala Arg Ala ArgAla Arg Ala Ala Leu Ala Arg Ala Ala Leu Ala Pro 50 55 60 Arg Leu Pro HisAsn Leu Ser Leu Glu Leu Val Val Ala Ala Pro Pro 65 70 75 80 Ala Arg AspPro Ala Ser Leu Thr Arg Gly Leu Cys Gln Ala Leu Val 85 90 95 Pro Pro GlyVal Ala Ala Leu Leu Ala Phe Pro Glu Ala Arg Pro Glu 100 105 110 Leu LeuGln Leu His Phe Leu Ala Ala Ala Thr Glu Thr Pro Val Leu 115 120 125 SerLeu Leu Arg Arg Glu Ala Arg Ala Pro Leu Gly Ala Pro Asn Pro 130 135 140Phe His Leu Gln Leu His Trp Ala Ser Pro Leu Glu Thr Leu Leu Asp 145 150155 160 Val Leu Val Ala Val Leu Gln Ala His Ala Trp Glu Asp Val Gly Leu165 170 175 Ala Leu Cys Arg Thr Gln Asp Pro Gly Gly Leu Val Ala Leu TrpThr 180 185 190 Ser Arg Ala Gly Arg Pro Pro Gln Leu Val Leu Asp Leu SerArg Arg 195 200 205 Asp Thr Gly Asp Ala Gly Leu Arg Ala Arg Leu Ala ProMet Ala Ala 210 215 220 Pro Val Gly Gly Glu Ala Pro Val Pro Ala Ala ValLeu Leu Gly Cys 225 230 235 240 Asp Ile Ala Arg Ala Arg Arg Val Leu GluAla Val Pro Pro Gly Pro 245 250 255 His Trp Leu Leu Gly Thr Pro Leu ProPro Lys Ala Leu Pro Thr Ala 260 265 270 Gly Leu Pro Pro Gly Leu Leu AlaLeu Gly Glu Val Ala Arg Pro Pro 275 280 285 Leu Glu Ala Ala Ile His AspIle Val Gln Leu Val Ala Arg Ala Leu 290 295 300 Gly Ser Ala Ala Gln ValGln Pro Lys Arg Ala Leu Leu Pro Ala Pro 305 310 315 320 Val Asn Cys GlyAsp Leu Gln Pro Ala Gly Pro Glu Ser Pro Gly Arg 325 330 335 Phe Leu AlaArg Phe Leu Ala Asn Thr Ser Phe Gln Gly Arg Thr Gly 340 345 350 Pro ValTrp Val Thr Gly Ser Ser Gln Val His Met Ser Arg His Phe 355 360 365 LysVal Trp Ser Leu Arg Arg Asp Pro Arg Gly Ala Pro Ala Trp Ala 370 375 380Thr Val Gly Ser Trp Arg Asp Gly Gln Leu Asp Leu Glu Pro Gly Gly 385 390395 400 Ala Ser Ala Arg Pro Pro Pro Pro Gln Gly Ala Gln Val Trp Pro Lys405 410 415 Leu Arg Val Val Thr Leu Leu Glu His Pro Phe Val Phe Ala ArgAsp 420 425 430 Pro Asp Glu Asp Gly Gln Cys Pro Ala Gly Gln Leu Cys LeuAsp Pro 435 440 445 Gly Thr Asn Asp Ser Ala Thr Leu Asp Ala Leu Phe AlaAla Leu Ala 450 455 460 Asn Gly Ser Ala Pro Arg Ala Leu Arg Lys Cys CysTyr Gly Tyr Cys 465 470 475 480 Ile Asp Leu Leu Glu Arg Leu Ala Glu AspThr Pro Phe Asp Phe Glu 485 490 495 Leu Tyr Leu Val Gly Asp Gly Lys TyrGly Ala Leu Arg Asp Gly Arg 500 505 510 Trp Thr Gly Leu Val Gly Asp LeuLeu Ala Gly Arg Ala His Met Ala 515 520 525 Val Thr Ser Phe Ser Ile AsnSer Ala Arg Ser Gln Val Val Asp Phe 530 535 540 Thr Ser Pro Phe Phe SerThr Ser Leu Gly Ile Met Val Arg Ala Arg 545 550 555 560 Asp Thr Ala SerPro Ile Gly Ala Phe Met Trp Pro Leu His Trp Ser 565 570 575 Thr Trp LeuGly Val Phe Ala Ala Leu His Leu Thr Ala Leu Phe Leu 580 585 590 Thr ValTyr Glu Trp Arg Ser Pro Tyr Gly Leu Thr Pro Arg Gly Arg 595 600 605 AsnArg Ser Thr Val Phe Ser Tyr Ser Ser Ala Leu Asn Leu Cys Tyr 610 615 620Ala Ile Leu Phe Arg Arg Thr Val Ser Ser Lys Thr Pro Lys Cys Pro 625 630635 640 Thr Gly Arg Leu Leu Met Asn Leu Trp Ala Ile Phe Cys Leu Leu Val645 650 655 Leu Ser Ser Tyr Thr Ala Asn Leu Ala Ala Val Met Val Gly AspLys 660 665 670 Thr Phe Glu Glu Leu Ser Gly Ile His Asp Pro Lys Leu HisHis Pro 675 680 685 Ala Gln Gly Phe Arg Phe Gly Thr Val Trp Glu Ser SerAla Glu Ala 690 695 700 Tyr Ile Lys Lys Ser Phe Pro Asp Met His Ala HisMet Arg Arg His 705 710 715 720 Ser Ala Pro Thr Thr Pro Arg Gly Val AlaMet Leu Thr Ser Asp Pro 725 730 735 Pro Lys Leu Asn Ala Phe Ile Met AspLys Ser Leu Leu Asp Tyr Glu 740 745 750 Val Ser Ile Asp Ala Asp Cys LysLeu Leu Thr Val Gly Lys Pro Phe 755 760 765 Ala Ile Glu Gly Tyr Gly IleGly Leu Pro Gln Asn Ser Pro Leu Thr 770 775 780 Ser Asn Leu Ser Glu PheIle Ser Arg Tyr Lys Ser Ser Gly Phe Ile 785 790 795 800 Asp Leu Leu HisAsp Lys Trp Tyr Lys Met Val Pro Cys Gly Lys Arg 805 810 815 Val Phe AlaVal Thr Glu Thr Leu Gln Met Ser Ile Tyr His Phe Ala 820 825 830 Gly LeuPhe Val Leu Leu Cys Leu Gly Leu Gly Ser Ala Leu Leu Ser 835 840 845 SerLeu Gly Glu His Ala Phe Phe Arg Leu Ala Leu Pro Arg Ile Arg 850 855 860Lys Gly Ser Arg Leu Gln Tyr Trp Leu His Thr Ser Gln Lys Ile His 865 870875 880 Arg Ala Leu Asn Thr Glu Pro Pro Glu Gly Ser Lys Glu Glu Thr Ala885 890 895 Glu Ala Glu Pro Arg 900 34 474 PRT Artificial SequenceConsensus Sequence 34 Xaa Val Xaa Xaa Xaa Xaa Pro Asp Glu Asp Gly GlnCys Pro Ala Gly 1 5 10 15 Gln Leu Cys Leu Asp Pro Gly Thr Asn Asp SerAla Thr Leu Asp Ala 20 25 30 Leu Phe Ala Ala Leu Ala Asn Gly Ser Ala ProArg Ala Leu Arg Lys 35 40 45 Cys Cys Tyr Gly Tyr Cys Ile Asp Leu Leu GluArg Leu Ala Glu Asp 50 55 60 Thr Pro Phe Asp Phe Glu Leu Tyr Leu Val GlyAsp Gly Lys Tyr Gly 65 70 75 80 Ala Leu Arg Asp Gly Arg Trp Thr Gly LeuVal Gly Asp Leu Leu Ala 85 90 95 Gly Arg Ala His Met Ala Val Thr Ser PheSer Ile Asn Ser Ala Arg 100 105 110 Ser Gln Val Val Asp Phe Thr Ser ProPhe Phe Ser Thr Ser Leu Gly 115 120 125 Ile Met Val Arg Ala Arg Asp ThrAla Ser Pro Ile Gly Ala Phe Met 130 135 140 Trp Pro Leu His Trp Ser ThrTrp Leu Gly Val Phe Ala Ala Leu His 145 150 155 160 Leu Thr Ala Leu PheLeu Thr Val Tyr Glu Trp Arg Ser Pro Tyr Gly 165 170 175 Leu Thr Pro ArgGly Arg Asn Arg Ser Thr Val Phe Ser Tyr Ser Ser 180 185 190 Ala Leu AsnLeu Cys Tyr Ala Ile Leu Phe Arg Arg Thr Val Ser Ser 195 200 205 Lys ThrPro Lys Cys Pro Thr Gly Arg Leu Leu Met Asn Leu Trp Ala 210 215 220 IlePhe Cys Leu Leu Val Leu Ser Ser Tyr Thr Ala Asn Leu Ala Ala 225 230 235240 Val Met Val Gly Asp Lys Thr Phe Glu Glu Leu Ser Gly Ile His Asp 245250 255 Pro Lys Xaa Xaa Xaa Xaa Xaa Xaa Gly Phe Arg Phe Gly Thr Val Trp260 265 270 Glu Ser Ser Ala Glu Ala Tyr Ile Lys Lys Ser Phe Pro Asp MetHis 275 280 285 Ala His Met Arg Arg His Ser Ala Pro Thr Thr Pro Arg GlyVal Ala 290 295 300 Met Leu Thr Ser Asp Pro Pro Lys Leu Asn Ala Phe IleMet Asp Lys 305 310 315 320 Ser Leu Leu Asp Tyr Glu Val Ser Ile Asp AlaAsp Cys Lys Leu Leu 325 330 335 Thr Val Gly Lys Pro Phe Ala Ile Glu GlyTyr Gly Ile Gly Leu Pro 340 345 350 Gln Asn Ser Pro Leu Thr Ser Asn LeuSer Glu Phe Ile Ser Arg Tyr 355 360 365 Lys Ser Ser Gly Phe Ile Asp LeuLeu His Asp Lys Trp Tyr Lys Met 370 375 380 Val Pro Cys Gly Lys Arg ValPhe Ala Val Thr Glu Thr Leu Gln Met 385 390 395 400 Ser Ile Tyr His PheAla Gly Leu Phe Val Leu Leu Cys Leu Gly Leu 405 410 415 Gly Ser Ala LeuLeu Ser Ser Leu Gly Glu His Ala Phe Phe Arg Leu 420 425 430 Ala Leu ProArg Ile Arg Lys Gly Ser Arg Leu Gln Tyr Trp Leu His 435 440 445 Thr SerGln Lys Ile His Arg Ala Leu Asn Thr Glu Pro Pro Glu Gly 450 455 460 SerLys Glu Glu Thr Ala Glu Ala Glu Pro 465 470 35 1135 PRT Rattusnorvegicus 35 Met Arg Arg Leu Ser Leu Trp Trp Leu Leu Ser Arg Val CysLeu Leu 1 5 10 15 Leu Pro Pro Pro Cys Ala Leu Val Leu Ala Gly Val ProSer Ser Ser 20 25 30 Ser His Pro Gln Pro Cys Gln Ile Leu Lys Arg Ile GlyHis Ala Val 35 40 45 Arg Val Gly Ala Val His Leu Gln Pro Trp Thr Thr AlaPro Arg Ala 50 55 60 Ala Ser Arg Ala Gln Glu Gly Gly Arg Ala Gly Ala GlnArg Asp Asp 65 70 75 80 Pro Glu Ser Gly Thr Trp Arg Pro Pro Ala Pro SerGln Gly Ala Arg 85 90 95 Trp Leu Gly Ser Ala Leu His Gly Arg Gly Pro ProGly Ser Arg Lys 100 105 110 Leu Gly Glu Gly Ala Gly Ala Glu Thr Leu TrpPro Arg Asp Ala Leu 115 120 125 Leu Phe Ala Val Glu Asn Leu Asn Arg ValGlu Gly Leu Leu Pro Tyr 130 135 140 Asn Leu Ser Leu Glu Val Val Met AlaIle Glu Ala Gly Leu Gly Asp 145 150 155 160 Leu Pro Leu Met Pro Phe SerSer Pro Ser Ser Pro Trp Ser Ser Asp 165 170 175 Pro Phe Ser Phe Leu GlnSer Val Cys His Thr Val Val Val Gln Gly 180 185 190 Val Ser Ala Leu LeuAla Phe Pro Gln Ser Gln Gly Glu Met Met Glu 195 200 205 Leu Asp Leu ValSer Ser Val Leu His Ile Pro Val Leu Ser Ile Val 210 215 220 Arg His GluPhe Pro Arg Glu Ser Gln Asn Pro Leu His Leu Gln Leu 225 230 235 240 SerLeu Glu Asn Ser Leu Ser Ser Asp Ala Asp Val Thr Val Ser Ile 245 250 255Leu Thr Met Asn Asn Trp Tyr Asn Phe Ser Leu Leu Leu Cys Gln Glu 260 265270 Asp Trp Asn Ile Thr Asp Phe Leu Leu Leu Thr Glu Asn Asn Ser Lys 275280 285 Phe His Leu Glu Ser Val Ile Asn Ile Thr Ala Asn Leu Ser Ser Thr290 295 300 Lys Asp Leu Leu Ser Phe Leu Gln Val Gln Met Asp Asn Ile ArgAsn 305 310 315 320 Ser Thr Pro Thr Met Val Met Phe Gly Cys Asp Met AspSer Ile Arg 325 330 335 Gln Ile Phe Glu Met Ser Thr Gln Phe Gly Leu SerPro Pro Glu Leu 340 345 350 His Trp Val Leu Gly Asp Ser Gln Asn Val GluGlu Leu Arg Thr Glu 355 360 365 Gly Leu Pro Leu Gly Leu Ile Ala His GlyLys Thr Thr Gln Ser Val 370 375 380 Phe Glu Tyr Tyr Val Gln Asp Ala MetGlu Leu Val Ala Arg Ala Val 385 390 395 400 Ala Thr Ala Thr Met Ile GlnPro Glu Leu Ala Leu Leu Pro Ser Thr 405 410 415 Met Asn Cys Met Asp ValLys Thr Thr Asn Leu Thr Ser Gly Gln Tyr 420 425 430 Leu Ser Arg Phe LeuAla Asn Thr Thr Phe Arg Gly Leu Ser Gly Ser 435 440 445 Ile Lys Val LysGly Ser Thr Ile Ile Ser Ser Glu Asn Asn Phe Phe 450 455 460 Ile Trp AsnLeu Gln His Asp Pro Met Gly Lys Pro Met Trp Thr Arg 465 470 475 480 LeuGly Ser Trp Gln Gly Gly Arg Ile Val Met Asp Ser Gly Ile Trp 485 490 495Pro Glu Gln Ala Gln Arg His Lys Thr His Phe Gln His Pro Asn Lys 500 505510 Leu His Leu Arg Val Val Thr Leu Ile Glu His Pro Phe Val Phe Thr 515520 525 Arg Glu Val Asp Asp Glu Gly Leu Cys Pro Ala Gly Gln Leu Cys Leu530 535 540 Asp Pro Met Thr Asn Asp Ser Ser Met Leu Asp Arg Leu Phe SerSer 545 550 555 560 Leu His Ser Ser Asn Asp Thr Val Pro Ile Lys Phe LysLys Cys Cys 565 570 575 Tyr Gly Tyr Cys Ile Asp Leu Leu Glu Gln Leu AlaGlu Asp Met Asn 580 585 590 Phe Asp Phe Asp Leu Tyr Ile Val Gly Asp GlyLys Tyr Gly Ala Trp 595 600 605 Lys Asn Gly His Trp Thr Gly Leu Val GlyAsp Leu Leu Ser Gly Thr 610 615 620 Ala Asn Met Ala Val Thr Ser Phe SerIle Asn Thr Ala Arg Ser Gln 625 630 635 640 Val Ile Asp Phe Thr Ser ProPhe Phe Ser Thr Ser Leu Gly Ile Leu 645 650 655 Val Arg Thr Arg Asp ThrAla Ala Pro Ile Gly Ala Phe Met Trp Pro 660 665 670 Leu His Trp Thr MetTrp Leu Gly Ile Phe Val Ala Leu His Ile Thr 675 680 685 Ala Ile Phe LeuThr Leu Tyr Glu Trp Lys Ser Pro Phe Gly Met Thr 690 695 700 Pro Lys GlyArg Asn Arg Asn Lys Val Phe Ser Phe Ser Ser Ala Leu 705 710 715 720 AsnVal Cys Tyr Ala Leu Leu Phe Gly Arg Thr Ala Ala Ile Lys Pro 725 730 735Pro Lys Cys Trp Thr Gly Arg Phe Leu Met Asn Leu Trp Ala Ile Phe 740 745750 Cys Met Phe Cys Leu Ser Thr Tyr Thr Ala Asn Leu Ala Ala Val Met 755760 765 Val Gly Glu Lys Ile Tyr Glu Glu Leu Ser Gly Ile His Asp Pro Lys770 775 780 Leu His His Pro Ser Gln Gly Phe Arg Phe Gly Thr Val Arg GluSer 785 790 795 800 Ser Ala Glu Asp Tyr Val Arg Gln Ser Phe Pro Glu MetHis Glu Tyr 805 810 815 Met Arg Arg Tyr Asn Val Pro Ala Thr Pro Asp GlyVal Gln Tyr Leu 820 825 830 Lys Asn Asp Pro Glu Lys Leu Asp Ala Phe IleMet Asp Lys Ala Leu 835 840 845 Leu Asp Tyr Glu Val Ser Ile Asp Ala AspCys Lys Leu Leu Thr Val 850 855 860 Gly Lys Pro Phe Ala Ile Glu Gly TyrGly Ile Gly Leu Pro Pro Asn 865 870 875 880 Ser Pro Leu Thr Ser Asn IleSer Glu Leu Ile Ser Gln Tyr Lys Ser 885 890 895 His Gly Phe Met Asp ValLeu His Asp Lys Trp Tyr Lys Val Val Pro 900 905 910 Cys Gly Lys Arg SerPhe Ala Val Thr Glu Thr Leu Gln Met Gly Ile 915 920 925 Lys His Phe SerGly Leu Phe Val Leu Leu Cys Ile Gly Phe Gly Leu 930 935 940 Ser Ile LeuThr Thr Ile Gly Glu His Ile Val His Arg Leu Leu Leu 945 950 955 960 ProArg Ile Lys Asn Lys Ser Lys Leu Gln Tyr Trp Leu His Thr Ser 965 970 975Gln Arg Phe His Arg Ala Leu Asn Thr Ser Phe Val Glu Glu Lys Gln 980 985990 Pro Arg Ser Lys Thr Lys Arg Val Glu Lys Ser Arg Trp Arg Arg Trp 9951000 1005 Thr Cys Lys Thr Glu Gly Asp Ser Glu Leu Ser Leu Phe Pro ArgSer 1010 1015 1020 Asn Leu Gly Pro Gln Gln Leu Met Val Trp Asn Thr SerAsn Leu Ser 1025 1030 1035 1040 His Asp Asn Gln Arg Lys Tyr Ile Phe AsnAsp Glu Glu Gly Gln Asn 1045 1050 1055 Gln Leu Gly Thr Gln Ala His GlnAsp Ile Pro Leu Pro Gln Arg Arg 1060 1065 1070 Arg Glu Leu Pro Ala SerLeu Thr Thr Asn Gly Lys Ala Asp Ser Leu 1075 1080 1085 Asn Val Thr ArgSer Ser Val Ile Gln Glu Leu Ser Glu Leu Glu Lys 1090 1095 1100 Gln IleGln Val Ile Arg Gln Glu Leu Gln Leu Ala Val Ser Arg Lys 1105 1110 11151120 Thr Glu Leu Glu Glu Tyr Gln Lys Thr Asn Arg Thr Cys Glu Ser 11251130 1135 36 1120 DNA Artificial Sequence Consensus Sequence 36cnctgccncc nangnncctg nccancncgn nnctnccann nnngcnncng gcncngngng 60nggtgncang ancnccncng gnanncnngn cncnncncnt nannntgtgn ancnnntngc 120cnggncncnn gggnngnncn gnccnggnnc ancngnngnn agcnnncnnn ncngccnngn 180tnnactnnng gnaccngnag nngncnnngc ncgngncccc gnnncnncnn nntgncnngg 240tnnctggccn aancnncgtn ntnnnanngc ngnnngnncn cccntntgng tnnnngncng 300nnnnccagac gaagacgggc agtgcccagc ggggcagctg tgcctggacc ctggcaccaa 360cgactcggcc accctggacg cactgttcgc cgcgctggcc aacggctcag cgccccgtgc 420cctgcgcaag tgctgctacg gctactgcat tgacctgctg gagcggctgg cggaggacac 480gcccttcgac ttcgagctgt acctcgtggg tgacggcaag tacggcgccc tgcgggacgg 540ccgctggacc ggcctggtcg gggacctgct ggccggccgg gcccacatgg cggtcaccag 600cttcagtatc aactccgccc gctcacaggt ggtggacttc accagcccct tcttctccac 660cagcctgggc atcatggtgc gggcacggga cacggcctca cccatcggtg cctttatgtg 720gcccctgcac tggtccacgt ggctgggcgt ctttgcggcc ctgcacctca ccgcgctctt 780cctcaccgtg tacgagtggc gtagccccta cggcctcacg ccacgtggcc gcaaccgcag 840caccgtcttc tcctactcct cagccctcaa cctgtgctac gccatcctct tcagacgcac 900cgtgtccagc aagacgccca agtgccccac gggccgcctg ctcatgaacc tctgggccat 960cttctgcctg ctggtgctgt ccagctacac ggccaacctg gctgccgtca tggtcgggga 1020caagaccttc gaggagctgt cggggatcca cgaccccaag ntgnncnncc ncggngnngg 1080gctncngntn nggcnnngng ngggnnagcn gngnccnngg 1120 37 474 PRT ArtificialSequence Consensus Sequence 37 Xaa Val Xaa Xaa Xaa Xaa Pro Asp Glu AspGly Gln Cys Pro Ala Gly 1 5 10 15 Gln Leu Cys Leu Asp Pro Gly Thr AsnAsp Ser Ala Thr Leu Asp Ala 20 25 30 Leu Phe Ala Ala Leu Ala Asn Gly SerAla Pro Arg Ala Leu Arg Lys 35 40 45 Cys Cys Tyr Gly Tyr Cys Ile Asp LeuLeu Glu Arg Leu Ala Glu Asp 50 55 60 Thr Pro Phe Asp Phe Glu Leu Tyr LeuVal Gly Asp Gly Lys Tyr Gly 65 70 75 80 Ala Leu Arg Asp Gly Arg Trp ThrGly Leu Val Gly Asp Leu Leu Ala 85 90 95 Gly Arg Ala His Met Ala Val ThrSer Phe Ser Ile Asn Ser Ala Arg 100 105 110 Ser Gln Val Val Asp Phe ThrSer Pro Phe Phe Ser Thr Ser Leu Gly 115 120 125 Ile Met Val Arg Ala ArgAsp Thr Ala Ser Pro Ile Gly Ala Phe Met 130 135 140 Trp Pro Leu His TrpSer Thr Trp Leu Gly Val Phe Ala Ala Leu His 145 150 155 160 Leu Thr AlaLeu Phe Leu Thr Val Tyr Glu Trp Arg Ser Pro Tyr Gly 165 170 175 Leu ThrPro Arg Gly Arg Asn Arg Ser Thr Val Phe Ser Tyr Ser Ser 180 185 190 AlaLeu Asn Leu Cys Tyr Ala Ile Leu Phe Arg Arg Thr Val Ser Ser 195 200 205Lys Thr Pro Lys Cys Pro Thr Gly Arg Leu Leu Met Asn Leu Trp Ala 210 215220 Ile Phe Cys Leu Leu Val Leu Ser Ser Tyr Thr Ala Asn Leu Ala Ala 225230 235 240 Val Met Val Gly Asp Lys Thr Phe Glu Glu Leu Ser Gly Ile HisAsp 245 250 255 Pro Lys Leu His His Pro Ala Gln Gly Phe Arg Phe Gly ThrVal Trp 260 265 270 Glu Ser Ser Ala Glu Ala Tyr Ile Lys Lys Ser Phe ProAsp Met His 275 280 285 Ala His Met Arg Arg His Ser Ala Pro Thr Thr ProArg Gly Val Ala 290 295 300 Met Leu Thr Ser Asp Pro Pro Lys Leu Asn AlaPhe Ile Met Asp Lys 305 310 315 320 Ser Leu Leu Asp Tyr Glu Val Ser IleAsp Ala Asp Cys Lys Leu Leu 325 330 335 Thr Val Gly Lys Pro Phe Ala IleGlu Gly Tyr Gly Ile Gly Leu Pro 340 345 350 Gln Asn Ser Pro Leu Thr SerAsn Leu Ser Glu Phe Ile Ser Arg Tyr 355 360 365 Lys Ser Ser Gly Phe IleAsp Leu Leu His Asp Lys Trp Tyr Lys Met 370 375 380 Val Pro Cys Gly LysArg Val Phe Ala Val Thr Glu Thr Leu Gln Met 385 390 395 400 Ser Ile TyrHis Phe Ala Gly Leu Phe Val Leu Leu Cys Leu Gly Leu 405 410 415 Gly SerAla Leu Leu Ser Ser Leu Gly Glu His Ala Phe Phe Arg Leu 420 425 430 AlaLeu Pro Arg Ile Arg Lys Gly Ser Arg Leu Gln Tyr Trp Leu His 435 440 445Thr Ser Gln Lys Ile His Arg Ala Leu Asn Thr Glu Pro Pro Glu Gly 450 455460 Ser Lys Glu Glu Thr Ala Glu Ala Glu Pro 465 470 38 1094 DNAArtificial Sequence Consensus Sequence 38 ggttcctggc caacacgtccttccagggcc gcacgggccc cgtgtgggtg acaggcagct 60 cccaggtaca catgtctcggcactttaagg tgtggagcct tcgccgggac ccacggggcg 120 ccccggcctg ggccacggtgggcagctggc gggacggcca gctggacttg gaaccgggag 180 gtgcctctgc acggcccccgcccccacagg gtgcccaggt ctggcccaag ctgcgtgtgg 240 taacgctgtt ggaacacccatttgtgtttg cccgtgatcc agacgaagac gggcagtgcc 300 cagcggggca gctgtgcctggaccctggca ccaacgactc ggccaccctg gacgcactgt 360 tcgccgcgct ggccaacggctcagcgcccc gtgccctgcg caagtgctgc tacggctact 420 gcattgacct gctggagcggctggcggagg acacgccctt cgacttcgag ctgtacctcg 480 tgggtgacgg caagtacggcgccctgcggg acggccgctg gaccggcctg gtcggggacc 540 tgctggccgg ccgggcccacatggcggtca ccagcttcag tatcaactcc gcccgctcac 600 aggtggtgga cttcaccagccccttcttct ccaccagcct gggcatcatg gtgcgggcac 660 gggacacggc ctcacccatcggtgccttta tgtggcccct gcactggtcc acgtggctgg 720 gcgtctttgc ggccctgcacctcaccgcgc tcttcctcac cgtgtacgag tggcgtagcc 780 cctacggcct cacgccacgtggccgcaacc gcagcaccgt cttctcctac tcctcagccc 840 tcaacctgtg ctacgccatcctcttcagac gcaccgtgtc cagcaagacg cccaagtgcc 900 ccacgggccg cctgctcatgaacctctggg ccatcttctg cctgctggtg ctgtccagct 960 acacggccaa cctggctgccgtcatggtcg gggacaagac cttcgaggag ctgtcgggga 1020 tccacgaccc caagntgnncnnccncggng nngggctncn gntnnggcnn ngngngggnn 1080 agcngngncc nngg 1094 39286 PRT Caenorhabditis elegans 39 Arg Ser Thr Leu Val Asn Lys Glu ProAsp Ser Met Leu Ala His Met 1 5 10 15 Phe Lys Asp Lys Gly Val Trp GlyAsn Lys Gln Asp His Arg Gly Ala 20 25 30 Phe Leu Ile Asp Arg Ser Pro GluTyr Phe Glu Pro Ile Leu Asn Tyr 35 40 45 Leu Arg His Gly Gln Leu Ile ValAsn Asp Gly Ile Asn Leu Leu Gly 50 55 60 Val Leu Glu Glu Ala Arg Phe PheGly Ile Asp Ser Leu Ile Glu His 65 70 75 80 Leu Glu Val Ala Ile Lys AsnSer Gln Pro Pro Glu Asp His Ser Pro 85 90 95 Ile Ser Arg Lys Glu Phe ValArg Phe Leu Leu Ala Thr Pro Thr Lys 100 105 110 Ser Glu Leu Arg Cys GlnGly Leu Asn Phe Ser Gly Ala Asp Leu Ser 115 120 125 Arg Leu Asp Leu ArgTyr Ile Asn Phe Lys Met Ala Asn Leu Ser Arg 130 135 140 Cys Asn Leu AlaHis Ala Asn Leu Cys Cys Ala Asn Leu Glu Arg Ala 145 150 155 160 Asp LeuSer Gly Ser Val Leu Asp Cys Ala Asn Leu Gln Gly Val Lys 165 170 175 MetLeu Cys Ser Asn Ala Glu Gly Ala Ser Leu Lys Leu Cys Asn Phe 180 185 190Glu Asp Pro Ser Gly Leu Lys Ala Asn Leu Glu Gly Ala Asn Leu Lys 195 200205 Gly Val Asp Met Glu Gly Ser Gln Met Thr Gly Ile Asn Leu Arg Val 210215 220 Ala Thr Leu Lys Asn Ala Lys Leu Lys Asn Cys Asn Leu Arg Gly Ala225 230 235 240 Thr Leu Ala Gly Thr Asp Leu Glu Asn Cys Asp Leu Ser GlyCys Asp 245 250 255 Leu Gln Glu Ala Asn Leu Arg Gly Ser Asn Val Lys GlyAla Ile Phe 260 265 270 Glu Glu Met Leu Thr Pro Leu His Met Ser Gln SerVal Arg 275 280 285 40 903 DNA Rattus norvegicus 40 agacttctagcctgcccctc taacgtgatg gccgtggaca tagaatacag ctacagcagt 60 atggccccttctctgcgcag agagcgcttc accttcaaga tctcccccaa actgaacaag 120 ccactgaggccttgtattca gctgggcagc aaggatgaag ccggcagaat ggtggccccc 180 acagtacaggagaagaaggt gaagaagcgg gtgtccttcg ccgacaacca ggggctggcc 240 ctaacaatggtgaaagtgtt ctcggaattc gatgacccac tagatattcc gtttaacatc 300 actgagctcctagacaacat cgtgagtctg acgacagcag agagtgagag ctttgttttg 360 gattttccgcagccttctgc agattactta gactttagaa atcggcttca gaccaaccat 420 gtctgcctcgaaaactgcgt gctgaaggag aaagccatcg cgggcaccgt caaggtccag 480 aacctggcattcgagaaggt tgtgaagatc agcatgacat tcgatacctg gaaaagcttc 540 acagacttcccttgtcagta tgtgaaggac acttacgctg gttcagacag ggacacattc 600 tcctttgatatcagcctacc ggagaaaatc cagtcttatg aaagaatgga gttcgccgtg 660 tgctacgagtgtaacggcca gtcgtactgg gacagcaaca aaggcaaaaa ctacaggatc 720 accagggccgaactcagatc cacccaggga atgactgagc cgtacaatgg gccggatttt 780 ggaatctcttttgaccagtt cgggagccct cggtgttcct tcggcctgtt tccagagtgg 840 cctagttatctggggtatga aaagctgggg ccctattact agtgagttga ctgcagttga 900 cag 903 41906 DNA Artificial Sequence Consensus Sequence 41 agnnttctag cctgnncntctancnnnntg atggcngtgg acatnganta cagntacanc 60 ngnatggcnc cttcnntgcgcnnagagngn ttnnccttna agatctcncc naancnnanc 120 aanccactga ggccttgtattcagctgngc agcaagnatg aagccngnng aatggtggcc 180 ccnncngtnc aggagaagaaggtgaanaag cgggtgtcct tcgcngacaa ccaggggctg 240 gccctnacaa tggtnaaagtgttctcggaa ttcgatgacc cnctagatat nccnttnaac 300 atcacngagc tcctagacaacatngtgagn ntgacgacag cagagagnga gagctttgtt 360 ntggattttn cncagccntctgcagattac ttagacttta gaaatcgnct tcagnccnac 420 cangtctgcc tnganaactgngtgctnaag ganaangcca tngcnggcac ngtnaaggtn 480 cagaacctng cattngagaagnnngtgaan atnagnatga cnttcganac ctggaanagc 540 tncacagact tnccttgtcagtangtgaag gacacttang cnggttcaga cagggacacn 600 ttctccttng anatcagcntnccngagaan atncagtctt atgaaagaat ggagttngcn 660 gtgtnctacg agtgnaanggncagncgtac tgggacagca acanaggcaa naactanagg 720 atcancnggg cnganntnanatcnacccag ggaatgacnn agccnnacan tggnccggat 780 ttnggaatnt cntttgaccagttcggnagc cctcggtgtt cctnnggnct gtttccagag 840 tggccnagtt anntnggntatgaaaagctn gggccctant actagtgann nnnctgcagn 900 tgacag 906 42 284 PRTRattus norvegicus 42 Met Ala Val Asp Ile Glu Tyr Ser Tyr Ser Ser Met AlaPro Ser Leu 1 5 10 15 Arg Arg Glu Arg Phe Thr Phe Lys Ile Ser Pro LysLeu Asn Lys Pro 20 25 30 Leu Arg Pro Cys Ile Gln Leu Gly Ser Lys Asp GluAla Gly Arg Met 35 40 45 Val Ala Pro Thr Val Gln Glu Lys Lys Val Lys LysArg Val Ser Phe 50 55 60 Ala Asp Asn Gln Gly Leu Ala Leu Thr Met Val LysVal Phe Ser Glu 65 70 75 80 Phe Asp Asp Pro Leu Asp Ile Pro Phe Asn IleThr Glu Leu Leu Asp 85 90 95 Asn Ile Val Ser Leu Thr Thr Ala Glu Ser GluSer Phe Val Leu Asp 100 105 110 Phe Pro Gln Pro Ser Ala Asp Tyr Leu AspPhe Arg Asn Arg Leu Gln 115 120 125 Thr Asn His Val Cys Leu Glu Asn CysVal Leu Lys Glu Lys Ala Ile 130 135 140 Ala Gly Thr Val Lys Val Gln AsnLeu Ala Phe Glu Lys Val Val Lys 145 150 155 160 Ile Arg Met Thr Phe AspThr Trp Lys Ser Phe Thr Asp Phe Pro Cys 165 170 175 Gln Tyr Val Lys AspThr Tyr Ala Gly Ser Asp Arg Asp Thr Phe Ser 180 185 190 Phe Asp Ile SerLeu Pro Glu Lys Ile Gln Ser Tyr Glu Arg Met Glu 195 200 205 Phe Ala ValCys Tyr Glu Cys Asn Gly Gln Ser Tyr Trp Asp Ser Asn 210 215 220 Lys GlyLys Asn Tyr Arg Ile Thr Arg Ala Glu Leu Arg Ser Thr Gln 225 230 235 240Gly Met Thr Glu Pro Tyr Asn Gly Pro Asp Phe Gly Ile Ser Phe Asp 245 250255 Gln Phe Gly Ser Pro Arg Cys Ser Phe Gly Leu Phe Pro Glu Trp Pro 260265 270 Ser Tyr Leu Gly Tyr Glu Lys Leu Gly Pro Tyr Tyr 275 280 43 282PRT Artificial Sequence Consensus Sequence 43 Met Ala Val Asp Ile GluTyr Xaa Tyr Xaa Xaa Met Ala Pro Ser Leu 1 5 10 15 Arg Xaa Glu Arg PheXaa Phe Lys Ile Ser Pro Lys Xaa Lys Pro Leu 20 25 30 Arg Pro Cys Ile GlnLeu Xaa Ser Lys Xaa Glu Ala Xaa Xaa Met Val 35 40 45 Ala Pro Xaa Val GlnGlu Lys Lys Val Lys Lys Arg Val Ser Phe Ala 50 55 60 Asp Asn Gln Gly LeuAla Leu Thr Met Val Lys Val Phe Ser Glu Phe 65 70 75 80 Asp Asp Pro LeuAsp Xaa Pro Phe Asn Ile Thr Glu Leu Leu Asp Asn 85 90 95 Ile Val Ser LeuThr Thr Ala Glu Ser Glu Ser Phe Val Leu Asp Phe 100 105 110 Xaa Gln ProSer Ala Asp Tyr Leu Asp Phe Arg Asn Arg Leu Gln Xaa 115 120 125 His ValCys Leu Glu Asn Cys Val Leu Lys Xaa Lys Ala Ile Ala Gly 130 135 140 ThrVal Lys Val Gln Asn Leu Ala Phe Glu Lys Xaa Val Lys Ile Arg 145 150 155160 Met Thr Phe Asp Thr Trp Lys Ser Xaa Thr Asp Phe Pro Cys Gln Tyr 165170 175 Val Lys Asp Thr Tyr Ala Gly Ser Asp Arg Asp Thr Phe Ser Phe Asp180 185 190 Ile Ser Leu Pro Glu Lys Ile Gln Ser Tyr Glu Arg Met Glu PheAla 195 200 205 Val Xaa Tyr Glu Cys Asn Gly Gln Xaa Tyr Trp Asp Ser AsnXaa Gly 210 215 220 Lys Asn Tyr Arg Ile Xaa Arg Ala Glu Leu Xaa Ser ThrGln Gly Met 225 230 235 240 Thr Xaa Pro Xaa Xaa Gly Pro Asp Xaa Gly IleSer Phe Asp Gln Phe 245 250 255 Gly Ser Pro Arg Cys Ser Xaa Gly Leu PhePro Glu Trp Pro Ser Tyr 260 265 270 Leu Gly Tyr Glu Lys Leu Gly Pro TyrTyr 275 280 44 294 PRT Mus musculus 44 Met Ala Met Arg Ile Cys Leu AlaHis Ser Pro Pro Leu Lys Ser Phe 1 5 10 15 Leu Gly Pro Tyr Asn Gly PheGln Arg Arg Asn Phe Val Asn Lys Leu 20 25 30 Lys Pro Leu Lys Pro Cys LeuSer Val Lys Gln Glu Ala Lys Ser Gln 35 40 45 Ser Glu Trp Lys Ser Pro HisAsn Gln Ala Lys Lys Arg Val Val Phe 50 55 60 Ala Asp Ser Lys Gly Leu SerLeu Thr Ala Ile His Val Phe Ser Asp 65 70 75 80 Leu Pro Glu Glu Pro AlaTrp Asp Leu Gln Phe Asp Leu Leu Asp Leu 85 90 95 Asn Asp Ile Ser Ser SerLeu Lys Leu His Glu Glu Lys Asn Leu Val 100 105 110 Phe Asp Phe Pro GlnPro Ser Thr Asp Tyr Leu Ser Phe Arg Asp Arg 115 120 125 Phe Gln Lys AsnPhe Val Cys Leu Glu Asn Cys Ser Leu Glu Asp Arg 130 135 140 Thr Val ThrGly Thr Val Lys Val Lys Asn Val Ser Phe Glu Lys Lys 145 150 155 160 ValGln Val Arg Ile Thr Phe Asp Thr Trp Lys Thr Tyr Thr Asp Val 165 170 175Asp Cys Val Tyr Met Lys Asn Val Tyr Ser Ser Ser Asp Ser Asp Thr 180 185190 Phe Ser Phe Ala Ile Asp Leu Pro Arg Val Ile Pro Thr Glu Glu Lys 195200 205 Ile Glu Phe Cys Ile Ser Tyr His Ala Asn Gly Arg Ile Phe Trp Asp210 215 220 Asn Asn Glu Gly Gln Asn Tyr Arg Ile Val His Val Gln Trp LysPro 225 230 235 240 Asp Gly Val Gln Thr Gln Val Ala Pro Lys Asp Cys AlaPhe Gln Gln 245 250 255 Gly Pro Pro Lys Thr Glu Ile Glu Pro Thr Val PheGly Ser Pro Arg 260 265 270 Leu Ala Ser Gly Leu Phe Pro Glu Trp Gln SerTrp Gly Arg Val Glu 275 280 285 Asn Leu Thr Ser Tyr Arg 290 45 312 PRTHomo sapiens 45 Met Ile Gln Val Leu Asp Pro Arg Pro Leu Thr Ser Ser ValMet Pro 1 5 10 15 Val Asp Val Ala Met Arg Leu Cys Leu Ala His Ser ProPro Val Lys 20 25 30 Ser Phe Leu Gly Pro Tyr Asp Glu Phe Gln Arg Arg HisPhe Val Asn 35 40 45 Lys Leu Lys Pro Leu Lys Ser Cys Leu Asn Ile Lys HisLys Ala Lys 50 55 60 Ser Gln Asn Asp Trp Lys Cys Ser His Asn Gln Ala LysLys Arg Val 65 70 75 80 Val Phe Ala Asp Ser Lys Gly Leu Ser Leu Thr AlaIle His Val Phe 85 90 95 Ser Asp Leu Pro Glu Glu Pro Ala Trp Asp Leu GlnPhe Asp Leu Leu 100 105 110 Asp Leu Asn Asp Ile Ser Ser Ala Leu Lys HisHis Glu Glu Lys Asn 115 120 125 Leu Ile Leu Asp Phe Pro Gln Pro Ser ThrAsp Tyr Leu Ser Phe Arg 130 135 140 Ser His Phe Gln Lys Asn Phe Val CysLeu Glu Asn Cys Ser Leu Gln 145 150 155 160 Glu Arg Thr Val Thr Gly ThrVal Lys Val Lys Asn Val Ser Phe Glu 165 170 175 Lys Lys Val Gln Ile ArgIle Thr Phe Asp Ser Trp Lys Asn Tyr Thr 180 185 190 Asp Val Asp Cys ValTyr Met Lys Asn Val Tyr Gly Gly Thr Asp Ser 195 200 205 Asp Thr Phe SerPhe Ala Ile Asp Leu Pro Pro Val Ile Pro Thr Glu 210 215 220 Gln Lys IleGlu Phe Cys Ile Ser Tyr His Ala Asn Gly Gln Val Phe 225 230 235 240 TrpAsp Asn Asn Asp Gly Gln Asn Tyr Arg Ile Val His Val Gln Trp 245 250 255Lys Pro Asp Gly Val Gln Thr Gln Met Ala Pro Gln Asp Cys Ala Phe 260 265270 His Gln Thr Ser Pro Lys Thr Glu Leu Glu Ser Thr Ile Phe Gly Ser 275280 285 Pro Arg Leu Ala Ser Gly Leu Phe Pro Glu Trp Gln Ser Trp Gly Arg290 295 300 Met Glu Asn Leu Ala Ser Tyr Arg 305 310 46 70 PRT Homosapiens 46 Met Pro Pro Asn Leu Thr Gly Tyr Tyr Arg Phe Val Ser Gln LysAsn 1 5 10 15 Met Glu Asp Tyr Leu Gln Ala Leu Asn Ile Ser Leu Ala ValArg Lys 20 25 30 Ile Ala Leu Leu Leu Lys Pro Asp Lys Glu Ile Glu His GlnGly Asn 35 40 45 His Met Thr Val Arg Thr Leu Ser Thr Phe Arg Asn Tyr ThrVal Gln 50 55 60 Phe Asp Val Gly Val Gln 65 70 47 70 PRT ArtificialSequence Consensus Sequence 47 Met Pro Pro Asn Leu Thr Gly Tyr Tyr ArgPhe Val Ser Gln Lys Asn 1 5 10 15 Met Glu Asp Tyr Leu Gln Ala Leu AsnIle Ser Leu Ala Val Arg Lys 20 25 30 Ile Ala Leu Leu Leu Lys Pro Asp LysGlu Ile Glu His Gln Gly Asn 35 40 45 His Met Thr Val Arg Thr Leu Ser ThrPhe Arg Asn Tyr Thr Xaa Gln 50 55 60 Phe Asp Val Gly Val Xaa 65 70 48135 PRT Homo sapiens 48 Met Pro Pro Asn Leu Thr Gly Tyr Tyr Arg Phe ValSer Gln Lys Asn 1 5 10 15 Met Glu Asp Tyr Leu Gln Ala Leu Asn Ile SerLeu Ala Val Arg Lys 20 25 30 Ile Ala Leu Leu Leu Lys Pro Asp Lys Glu IleGlu His Gln Gly Asn 35 40 45 His Met Thr Val Arg Thr Leu Ser Thr Phe ArgAsn Tyr Thr Leu Gln 50 55 60 Phe Asp Val Gly Val Glu Phe Glu Glu Asp LeuArg Ser Val Asp Gly 65 70 75 80 Arg Lys Cys Gln Thr Ile Val Thr Trp GluGlu Glu His Leu Val Cys 85 90 95 Val Gln Lys Gly Glu Val Pro Asn Arg GlyTrp Arg His Trp Leu Glu 100 105 110 Gly Glu Met Leu Tyr Leu Glu Leu ThrAla Arg Asp Ala Val Cys Glu 115 120 125 Gln Val Phe Arg Lys Val Arg 130135 49 135 PRT Artificial Sequence Consensus Sequence 49 Met Pro Pro AsnLeu Thr Gly Tyr Tyr Arg Phe Val Ser Gln Lys Asn 1 5 10 15 Met Glu AspTyr Leu Gln Ala Leu Asn Ile Ser Leu Ala Val Arg Lys 20 25 30 Ile Ala LeuLeu Leu Lys Pro Asp Lys Glu Ile Glu His Gln Gly Asn 35 40 45 His Met ThrVal Arg Thr Leu Ser Thr Phe Arg Asn Tyr Thr Xaa Gln 50 55 60 Phe Asp ValGly Val Glu Phe Glu Glu Asp Leu Arg Ser Val Asp Gly 65 70 75 80 Arg LysCys Gln Thr Ile Val Thr Trp Glu Glu Glu His Leu Val Cys 85 90 95 Val GlnLys Gly Glu Val Pro Asn Arg Gly Trp Arg His Trp Leu Glu 100 105 110 GlyGlu Xaa Leu Tyr Leu Glu Leu Thr Ala Arg Asp Ala Val Cys Glu 115 120 125Gln Val Phe Arg Lys Val Arg 130 135

What is claimed is:
 1. An isolated polypeptide comprising an amino acidsequence selected from the group consisting of: (a) a mature form of theamino acid sequence selected from the group consisting of SEQ ID NO:2,4, 6, 8, 10, 12, 14, or 16; (b) a variant of a mature form of the aminoacid sequence selected from the group consisting of SEQ ID NO:2, 4, 6,8, 10, 12, 14, or 16, wherein any amino acid in the mature form ischanged to a different amino acid, provided that no more than 20% of theamino acid residues in the sequence of the mature form are so changed;(c) the amino acid sequence selected from the group consisting of SEQ IDNO:2, 4, 6, 8, 10, 12, 14, or 16; (d) a variant of the amino acidsequence selected from the group consisting of SEQ ID NO:2, 4, 6, 8, 10,12, 14, or 16, wherein any amino acid specified in the chosen sequenceis changed to a different amino acid, provided that no more than 20% ofthe amino acid residues in the sequence are so changed; and (e) afragment of any of (a) through (d).
 2. The polypeptide of claim 1,wherein said polypeptide is a naturally occurring allelic variant of thesequence selected from the group consisting of SEQ ID NO:2, 4, 6, 8, 10,12, 14, or
 16. 3. The polypeptide of claim 2, wherein the allelicvariant is the translation of a single nucleotide polymorphism (SNP). 4.The polypeptide of claim 1 that is a variant polypeptide describedtherein, wherein any amino acid specified in the chosen sequence ischanged to provide a conservative substitution.
 5. An isolated nucleicacid molecule comprising a nucleic acid sequence encoding a polypeptidecomprising an amino acid sequence selected from the group consisting of:(a) a mature form of the amino acid sequence selected from the groupconsisting of SEQ ID NO:2, 4, 6, 8, 10, 12, 14, or 16; (b) a variant ofa mature form of the amino acid sequence selected from the groupconsisting of SEQ ID NO:2, 4, 6, 8, 10, 12, 14, or 16, wherein any aminoacid in the mature form of the chosen sequence is changed to a differentamino acid, provided that no more than 20% of the amino acid residues inthe sequence of the mature form are so changed; (c) the amino acidsequence selected from the group consisting of SEQ ID NO:2, 4,6,8,10,12, 14,or 16; (d) a variant of the amino acid sequence selected from thegroup consisting of SEQ ID NO:2, 4, 6, 8, 10, 12, 14, or 16, in whichany amino acid specified in the chosen sequence is changed to adifferent amino acid, provided that no more than 20% of the amino acidresidues in the sequence are so changed; (e) a nucleic acid fragmentencoding at least a portion of a polypeptide comprising the amino acidsequence selected from the group consisting of SEQ ID NO:2, 4, 6, 8, 10,12, 14, or 16, or any variant of said polypeptide wherein any amino acidof the chosen sequence is changed to a different amino acid, providedthat no more than 10% of the amino acid residues in the sequence are sochanged; and (f) the complement of any of said nucleic acid molecules.6. The nucleic acid molecule of claim 5, wherein the nucleic acidmolecule comprises the nucleotide sequence of a naturally occurringallelic nucleic acid variant.
 7. The nucleic acid molecule of claim 5that encodes a variant polypeptide, wherein the variant polypeptide hasthe polypeptide sequence of a naturally occurring polypeptide variant.8. The nucleic acid molecule of claim 5, wherein the nucleic acidmolecule comprises a single nucleotide polymorphism encoding saidvariant polypeptide.
 9. The nucleic acid molecule of claim 5, whereinsaid nucleic acid molecule comprises a nucleotide sequence selected fromthe group consisting of (a) the nucleotide sequence selected from thegroup consisting of SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, or 15; (b) anucleotide sequence wherein one or more nucleotides in the nucleotidesequence selected from the group consisting of SEQ ID NO: 1, 3, 5, 7, 9,11, 13, or 15 is changed from that given by the chosen sequence to adifferent nucleotide provided that no more than 20% of the nucleotidesare so changed; (c) a nucleic acid fragment of the sequence selectedfrom the group consisting of SEQ ID NO:1, 3,5,7,9, 11, 13, or 15; and(d) a nucleic acid fragment wherein one or more nucleotides in thenucleotide sequence selected from the group consisting of SEQ ID NO:1,3, 5, 7, 9, 11, 13, or 15 is changed from that given by the chosensequence to a different nucleotide provided that no more than 20% of thenucleotides are so changed.
 10. The nucleic acid molecule of claim 5,wherein said nucleic acid molecule hybridizes under stringent conditionsto the nucleotide sequence selected from the group consisting of SEQ IDNO:1, 3, 5, 7, 9, 11, 13, or 15, or a complement of said nucleotidesequence.
 11. The nucleic acid molecule of claim 5, wherein the nucleicacid molecule comprises a nucleotide sequence in which any nucleotidespecified in the coding sequence of the chosen nucleotide sequence ischanged from that given by the chosen sequence to a different nucleotideprovided that no more than 20% of the nucleotides in the chosen codingsequence are so changed, an isolated second polynucleotide that is acomplement of the first polynucleotide, or a fragment of any of them.12. A vector comprising the nucleic acid molecule of claim
 11. 13. Thevector of claim 12, further comprising a promoter operably linked tosaid nucleic acid molecule.
 14. A cell comprising the vector of claim12.
 15. An antibody that binds immunospecifically to the polypeptide ofclaim
 1. 16. The antibody of claim 15, wherein said antibody is amonoclonal antibody.
 17. The antibody of claim 15, wherein the antibodyis a humanized antibody.
 18. A method for determining the presence oramount of the polypeptide of claim 1 in a sample, the method comprising:(a) providing said sample; (b) introducing said sample to an antibodythat binds immunospecifically to the polypeptide; and (c) determiningthe presence or amount of antibody bound to said polypeptide, therebydetermining the presence or amount of polypeptide in said sample.
 19. Amethod for determining the presence or amount of the nucleic acidmolecule of claim 5 in a sample, the method comprising: (a) providingsaid sample; (b) introducing said sample to a probe that binds to saidnucleic acid molecule; and (c) determining the presence or amount ofsaid probe bound to said nucleic acid molecule, thereby determining thepresence or amount of the nucleic acid molecule in said sample.
 20. Amethod of identifying an agent that binds to the polypeptide of claim 1,the method comprising: (a) introducing said polypeptide to said agent;and (b) determining whether said agent binds to said polypeptide.
 21. Amethod for identifying a potential therapeutic agent for use intreatment of a pathology, wherein the pathology is related to aberrantexpression or aberrant physiological interactions of the polypeptide ofclaim 1, the method comprising: (a) providing a cell expressing thepolypeptide of claim 1 and having a property or function ascribable tothe polypeptide; (b) contacting the cell with a composition comprising acandidate substance; and determining whether the substance alters theproperty or function ascribable to the polypeptide; whereby, if analteration observed in the presence of the substance is not observedwhen the cell is contacted with a composition devoid of the substance,the substance is identified as a potential therapeutic agent.
 22. Amethod for modulating the activity of the polypeptide of claim 1, themethod comprising introducing a cell sample expressing the polypeptideof said claim with a compound that binds to said polypeptide in anamount sufficient to modulate the activity of the polypeptide.
 23. Amethod of treating or preventing a pathology associated with thepolypeptide of claim 1, said method comprising administering thepolypeptide of claim 1 to a subject in which such treatment orprevention is desired in an amount sufficient to treat or prevent saidpathology in said subject.
 24. The method of claim 23, wherein saidsubject is a human.
 25. A method of treating or preventing a pathologyassociated with the polypeptide of claim 1, said method comprisingadministering to a subject in which such treatment or prevention isdesired a MEMX nucleic acid in an amount sufficient to treat or preventsaid pathology in said subject.
 26. The method of claim 25, wherein saidsubject is a human.
 27. A method of treating or preventing a pathologyassociated with the polypeptide of claim 1, said method comprisingadministering to a subject in which such treatment or prevention isdesired a MEMX antibody in an amount sufficient to treat or prevent saidpathology in said subject.
 28. The method of claim 15, wherein thesubject is a human.
 29. A pharmaceutical composition comprising thepolypeptide of claim 1 and a pharmaceutically acceptable carrier.
 30. Apharmaceutical composition comprising the nucleic acid molecule of claim5 and a pharmaceutically acceptable carrier.
 31. A pharmaceuticalcomposition comprising the antibody of claim 15 and a pharmaceuticallyacceptable carrier.
 32. A kit comprising in one or more containers, thepharmaceutical composition of claim
 29. 33. A kit comprising in one ormore containers, the pharmaceutical composition of claim
 30. 34. A kitcomprising in one or more containers, the pharmaceutical composition ofclaim
 31. 35. The use of a therapeutic in the manufacture of amedicament for treating a syndrome associated with a human disease, thedisease selected from a pathology associated with the polypeptide ofclaim 1, wherein said therapeutic is the polypeptide of claim
 1. 36. Theuse of a therapeutic in the manufacture of a medicament for treating asyndrome associated with a human disease, the disease selected from apathology associated with the polypeptide of claim 1, wherein saidtherapeutic is a MEMX nucleic acid.
 37. The use of a therapeutic in themanufacture of a medicament for treating a syndrome associated with ahuman disease, the disease selected from a pathology associated with thepolypeptide of claim 1, wherein said therapeutic is a MEMX antibody. 38.A method for screening for a modulator of activity or of latency orpredisposition to a pathology associated with the polypeptide of claim1, said method comprising: (a) administering a test compound to a testanimal at increased risk for a pathology associated with the polypeptideof claim 1, wherein said test animal recombinantly expresses thepolypeptide of claim 1; (b) measuring the activity of said polypeptidein said test animal after administering the compound of step (a); and(c) comparing the activity of said protein in said test animal with theactivity of said polypeptide in a control animal not administered saidpolypeptide, wherein a change in the activity of said polypeptide insaid test animal relative to said control animal indicates the testcompound is a modulator of latency of, or predisposition to, a pathologyassociated with the polypeptide of claim
 1. 39. The method of claim 38,wherein said test animal is a recombinant test animal that expresses atest protein transgene or expresses said transgene under the control ofa promoter at an increased level relative to a wild-type test animal,and wherein said promoter is not the native gene promoter of saidtransgene.
 40. A method for determining the presence of orpredisposition to a disease associated with altered levels of thepolypeptide of claim 1 in a first mammalian subject, the methodcomprising: (a) measuring the level of expression of the polypeptide ina sample from the first mammalian subject; and (b) comparing the amountof said polypeptide in the sample of step (a) to the amount of thepolypeptide present in a control sample from a second mammalian subjectknown not to have, or not to be predisposed to, said disease, wherein analteration in the expression level of the polypeptide in the firstsubject as compared to the control sample indicates the presence of orpredisposition to said disease.
 41. A method for determining thepresence of or predisposition to a disease associated with alteredlevels of the nucleic acid molecule of claim 5 in a first mammaliansubject, the method comprising: (a) measuring the amount of the nucleicacid in a sample from the first mammalian subject; and (b) comparing theamount of said nucleic acid in the sample of step (a) to the amount ofthe nucleic acid present in a control sample from a second mammaliansubject known not to have or not be predisposed to, the disease; whereinan alteration in the level of the nucleic acid in the first subject ascompared to the control sample indicates the presence of orpredisposition to the disease.
 42. A method of treating a pathologicalstate in a mammal, the method comprising administering to the mammal apolypeptide in an amount that is sufficient to alleviate thepathological state, wherein the polypeptide is a polypeptide having anamino acid sequence at least 20% identical to a polypeptide comprisingthe amino acid sequence selected from the group consisting of SEQ IDNO:2, 4, 6, 8, 10, 12, 14, or 16, or a biologically active fragmentthereof.
 43. A method of treating a pathological state in a mammal, themethod comprising administering to the mammal the antibody of claim 15in an amount sufficient to alleviate the pathological state.